# Let's Freeze Some Yeast



## Bribie G

I recently started a Slanting thread as that's what I was intending to do, and got some great advice. However during the course of things, the alternative - freezing tubes of yeast - was presented so I had a good think about it and:







I have an old school freezer that needs de-icing every couple of months so the yeast tubes can be in deep freeze with no interruption as can happen with modern no frost cycling freezers. I acquired some kit and a bottle of glycerine (to prevent cell walls bursting which is the major no no in yeast freezing). I'll certainly revisit slanting if I get bad results down the track. Also please note I don't have a pressure cooker yet: relying on steaming only. 
So here goes:

8 test tubes (30ml) to be prepared for freezing. I'm following the method provided by this site: 

Into the stockpot: 
8 test tubes and caps sitting in a Pyrex bowl, 
a few assorted new syringes from the chemist sitting in a couple of Pyrex 250 ml teacups. 






We are looking to get a cup containing 120ml of 70% water / 30% glycerine, and a cup of 120ml yeast slurry to fill 8 tubes. 

Steam for an hour. 






Arrange a work area away from any spores and yeast as might lurk in your brewery. Using a small graduated syringe, put 85 ml of boiled kettle water and 35 ml glycerine into one of the glass cups or your equivalent bit of kit, and microwave to a boil. The into each test tube, syringe 15 ml of solution, cap the tubes and cool rapidly.
















Obtain a cup of slurry / yeast from a starter / topcrop from Yorkshire yeast etc etc.

Into each test tube, syringe 15 ml of yeasty solution. Cap, label and freeze.











Apparently if you fridge them for 24 hours and then freeze them you get better viability.


I look forward to the results. BTW the yeast is Wyeast 1968 London ESB. B)


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## Rodolphe01

Between all your experiments, do you get time to make beer that you will actually drink ?? 

I'll be watching this one with interest.


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## RdeVjun

Thanks Bribie for the great guide! B) 

There are other uses for those gloves after all!


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## rotten

Thanks Bribieg. 
Apart from actually just storing yeast cake or top cropping, that's the easiest and most uncomplicated explanation I have seen.
Cheers


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## Cocko

Legend.

:icon_cheers:


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## Eater

Immaculate
Look forward to seeing results


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## Adam Howard

What's involved once you thaw out the yeast to separate it from the glycerine Bribie?


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## white.grant

Hi Bribie

That's a good way of storing slurry for a longer period of time, though I don't think you should look to it as an alternative to slanting . 

The whole idea behind slanting is that you are using a fresh sample of yeast straight from the package and growing colonies in a controlled way so that they will not mutate. 

With a slant you can continue to re-culture and maintain the viability of your bank, but your slurry samples will have already mutated. 

cheers

grant


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## brettprevans

Interesting bribie. I remember Spillsmostofit freezing yeast years ago, I wonder what his method/results.

Freezing would be a good option to prevent yeast going off, just depends on how it affects it's viability


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## beerdrinkingbob

Hey Bribie,

Just a quick question to make sure i have it right, you rapid cool the mix say in the freezer till it goes solid?? and then add the yeast slurry.


Thanks again, great work much appreciated :super:


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## np1962

beerdrinkingbob said:


> Hey Bribie,
> 
> Just a quick question to make sure i have it right, you rapid cool the mix say in the freezer till it goes solid?? and then add the yeast slurry.
> 
> 
> Thanks again, great work much appreciated :super:


bob,
The idea is that the yeast combines/is surrounded by the glycerine solution which never quite freezes solid thereby holding the yeast in very low temperature conditions without doing damage to the cell walls.
Nige


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## schooey

Grantw said:


> Hi Bribie
> 
> That's a good way of storing slurry for a longer period of time, though I don't think you should look to it as an alternative to slanting .
> 
> The whole idea behind slanting is that you are using a fresh sample of yeast straight from the package and growing colonies in a controlled way so that they will not mutate.
> 
> With a slant you can continue to re-culture and maintain the viability of your bank, but your slurry samples will have already mutated.
> 
> cheers
> 
> grant



Great way to split a fresh smack pack into five and keep for an extended period though...


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## MeLoveBeer

Great work Bribie. Any chance of documenting the usage half of the process? Does the whole lot get pitched into your starter after being defrosted? (I can't imagine that the glycerine can be decanted)


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## Nevalicious

Me love beer

Follow this link

Post 7??

Bribie, I just did this last night too. Although I just used a smack pack, divided up into 4 glass 20ml vials with approx 5ml of glycerine in each, poured the remaining into a 700ml starter, and had it on my stir plate all night. Went right off!! :beerbang: Much like Schooey suggests above...

This may be a little dumb, but I checked my frozen samples this morning. Yes I know that they have been in the freezer, but they are rock solid. I thought the glycerine would keep them somewhat not frozen?? Anywho, time will tell when I go to step up from one of the samples. It'll either work or it wont hey!? 

When I use the starter in my next brew (this weekend) I might try my hand at top cropping. I have read Wyeast 1007 loves a good top cropping, and freeze some more. 

Nothing like making your dollar go further. 

Tyler


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## BjornJ

Great job documenting how to do it, thanks BribieG.

I store yeast in test tubes with sterile water over for months and years in the fridge, but this is probably a better way, eh.
And it doesn't look all that hard.

Hmm maybe something to try at some point.


If the vials freeze solid, did that mean the yeast is now dead and next time use more glycerin?

Bjorn


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## white.grant

schooey said:


> Great way to split a fresh smack pack into five and keep for an extended period though...



Perhaps, I guess it all depends on how viable the yeast are once they thaw. 

cheers

grant


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## Bribie G

I think the idea is that they do indeed freeze but the glycerine has stopped the cell walls from bursting, like freezing embryos etc. Should work ok with smackpacks - 125ml (if that's what is actually in a smack pack) should give 8 tubes as well, but would need a lot of culturing up afterwards compared to a whole smack pack. From what I've read the most popular method with a smack pack is to do a healthy starter to increase the yeast population, use the starter to do your tubes but have enough left over to pitch a brew in the normal manner. So with subsequent brews you are culturing up the same material that was in the original starter. 

Starting up and stepping up would be similar to slants but hopefully a bit quicker as it's a bigger yeast sample. Glycerine is quite edible and I guess the small amount that ends up in a brew would be negligible, but I would probably run up a starter to the point where there was a fair amount of yeast on the bottom, and pour off the upper liquid layer and then keep stepping up the starter with fresh wort until ready to pitch, would get rid of most of the glycerine.

Edit: last year I was clearing out the freezer to defrost it and there was a Wyeast (forget which) - had been sitting there for two months at least. It cultured up ok, although somewhat slower than usual. I reckon with the glycerine treatment it should work fine.


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## unrealeous

Nice work BribieG - but I'd suggest one slight change to your process for next time. That is - fill your containers with 85mls of glycerine, do the lid up tight and then pressure cook them. Once they are cooled - when you go to put the yeast slurry in - you only want the lids off for as short a time as possible. I say this since I used to do quite a bit of mushroom cultivation on agar slants - and anything uncovered for more than the shortest time would often end up with a rouge bacteria - and I see in your photos all the vials sitting there with the lids off after sterilization.

Anyway - good effort - love the photos - keen to see how well it works.


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## MeLoveBeer

Nevalicious said:


> Me love beer
> 
> Follow this link
> 
> Post 7??



Cheers Nevalicious. Have been splitting wyeast packs for a bit (just storing test tubes in the fridge) and am very interested in this as a long term storage solution.


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## argon

Great stuff Bribie, good simple guide to help everyone out. I'l be giving this a go shortly to keep my PC slants, as soon as i can source some tubes.

Cheers
:icon_cheers:


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## [email protected]

Bribie,

Good to see you having a go. It takes me months to get my crap together to do these things!

Just following through on Unrealeous. I would autoclave/pressure cook the tubes with the glycerin in them with the lids on and only take the lid off briefly to add yeast etc.

How are storing in your freezer as this is also critical to the process? Are using ice packs and a small esky to avoid the defrost cycle. I have purchased a few of the technice packs (50c each) to pack the vials in.

Maybe add a few photos of how you are storing to complete the OP.


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## drsmurto

Did you sterilise the glycerine?


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## Bribie G

I microwaved the glycerine / preboiled water mix in one of the pyrex cups as per the American Website and syringed immediately into the tubes then covered them.


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## drsmurto

BribieG said:


> I microwaved the glycerine / preboiled water mix in one of the pyrex cups as per the American Website and syringed immediately into the tubes then covered them.



All good, just didn't notice it in the steamer. :icon_cheers: 

Like you i want to do this without having to buy a pressure cooker.


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## drsmurto

Huge thanks bribie, just followed the link to another link on canning wort - Link

As it happens my HLT is actually a Fowlers Vacola preserving urn and i own several boxes of preserving jars as well as 100s of jam jars.

I have an excess of JW trad so making up 20L of wort at 1.040 and then preserving them for starters later sound much simpler than forking out money for extract.

I had done this in the past and frozen the wort in 1.5L juice/soft drink bottles but that takes up a lot of space in the freezer. Preserving them means they don't need to be frozen.

Cheers
DrSmurto

p.s any comments on not boiling the wort prior to putting it in the preserving jars? This would be the same as doing a boil with the lid on.


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## choppadoo

I work in a molecular biology lab, and we regularly make glycerol stocks for long term storage. Usually for bacterial cultures, but the principle is the same for yeast. I have just started making glycerol stocks of wyeasts that I buy. We store in -80C freezers, basically the colder you get it, the longer the cells keep their vitality, and the less often you have to re-culture. I make my glycerols with a final concentration of 25% glycerol, the other 75% being taken directly from yeast smackpacks. I make 1mL tubes, if you have good sterile procedures, you can make starter cultures from a single glycerol stock almost indefinitely (this will also reduce the amount of freezer space required). This you do by streaking a swab from the glycerol onto an agar plate, selecting a healthy colony and innoculating into progressively larger cultures (e.g. 3mL, 25mL, 250 mL, 1L, wort!). The agar plate step is probably not neccessary in a homebrew situation. This protocol will only work if you have good sterile technique; bacterial cultures are prepared in exactly the same way! Also, do not chill the glycerols slowly, reccomended best practice is to snap-freeze in liquid nitrogen before placing into the freezer, although this is not really neccesary.
If done correctly, glycerol stocks are a much better long term storage option for yeast than slants; they will retain vitality for much longer.

Cheers,
Brad


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## Nevalicious

BribieG said:


> I microwaved the glycerine / preboiled water mix in one of the pyrex cups as per the American Website and syringed immediately into the tubes then covered them.



Whoops, I used just 25% glycerine, and the rest was plain ol smacked Wyeast. Forgot the water...

I'm sure it will be fine though. Maybe <_<


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## argon

Froze me some yeast last night :beerbang: 

Tried it first with some good ol' US 05... i figure if things don't work out no loss. If it's all good i'll be freezing some precious 1469 i have in the fridge.

Couldn't have been easier... especially considering i used sample jars and syringes that are pre-sterilised in the packet.
Just made up my solution of Glycerin and pre-boiled water, brought it to boil again in microwave, added to sample jars, added appropriate amount (35ml in my case), then took some top cropped washed yeast and syringed 10ml each into 5 jars.

I now have 50ml of washed, frozen top crop US05 ready for starting up whenever i want. I can see this yeast harvesting and storage thing getting out of hand and filling up my freezer with sample jars of murky brown liquid. :blink: 
:icon_cheers:


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## davo4772

I'm not too fussed about using yeast over and over but i would like to reuse yeast at a later time one or twice to reduce costs.

I am able to access larger sterile containers. I wonder if you could freeze the required amount (for another batch) of yeast cake in one container.

Prior to brew day unthaw the yeast cake, add a pint or so of starter to rouse the yeast and Bob's your aunty.

Can't see why it would not work. Depends on loss of viability during storage. The minimum number of cells you put to sleep might not wake up.

My attraction to this method would be not having to step up more than once. 

Cheers


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## Bribie G

OK it's been nearly 4 months and the original tubes from the OP have been sitting in the back of the freezer - I've just cubed a London style Special Bitter and decided to thaw a couple of tubes and see how they go.

I collected a litre of end-of-runnings out of the urn into a Schott Bottle and let them settle out, and got about 300ml of clear stuff that I've poured into another 1L Schott bottle, and slid a couple of frozen plugs out of the test tubes, good shake, and it's now probably around 12 degrees, let it warm up and see how the cow sits in the ol' cabbage patch by tomorrow. :icon_cheers:


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## Punkal

Good luck Bribie... I hope they are trying to climb their way out the top of the schott bottle by tomorrow morning...


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## Yob

BribieG said:


> let it warm up and see how the cow sits in the ol' cabbage patch by tomorrow. :icon_cheers:



Hay Bribie.. just curious, Dave Line in "The Big Book of Brewing" talks about freezing a "Ball" gained by collecting Yeast from the Krausen (the middle section).. did you give that a go at any point?

I dont know when it was written but is this method no longer used?

Great pictorial too.. nice one

:icon_cheers:


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## Bribie G

Comrades
It is with deepest sorrow that I report that the brave and patriotic yeasties did not make it through Cryo.





I suspect my freezer just wasn't cold enough, well onto some slanting now and see if that works better - have the agar.

In recognition of their selfless sacrifice a Plaza has been renamed in their honour "La Plaza de Los Valientes Levaduras de La 18 Abril" and the Chairman will snip the ribbon this afternoon. :beerbang:


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## adryargument

Not good, i may need to test mine....


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## QldKev

BribieG said:


> Comrades
> It is with deepest sorrow that I report that the brave and patriotic yeasties did not make it through Cryo.
> 
> View attachment 45323
> 
> 
> I suspect my freezer just wasn't cold enough, well onto some slanting now and see if that works better - have the agar.
> 
> In recognition of their selfless sacrifice a Plaza has been renamed in their honour "La Plaza de Los Valientes Levaduras de La 18 Abril" and the Chairman will snip the ribbon this afternoon. :beerbang:



I wonder what the issue is with the yeast? I have yeast in test tubes dating back to June 2009, that has only been stored in the fridge. They are still happy yeasties again within 24-36 hours (CPA is sometimes up to 48hrs) getting back to doing their rude stuff in my wort. I would have though a freezer even at the higher end of the freezing range would have been plently cold, when my fridge is set to 6.0c. 

QldKev


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## Bribie G

Yes it's a mystery. I've had to move on quickly with the current brew so I'm using some 1469 cake instead. I think I'll do up a LDME wort, pitch another couple of tubes and see if they do kick off after a few days. However no movement at all over the last couple of days, and I'd expect something.


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## ratchie

Are you letting the yeast thaw out for a couple of days before pitching.


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## MHB

I think you didnt have enough Glycerine, from my understanding you need enough to act as an anti freeze.

When I was mucking around with this a couple of years ago I put the yeast slurry into tubes, let it settle overnight, removed the superannuant liquid (beer) then topped up with 30% Glycerine solution, target being 25% Glycerine and there is some argument that up to 35% might be better.

Your Glycerine content is less than 15%.

Be worth trying again, I got good viability up to 2 years out.
MHB


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## QldKev

What MHB said makes sense. If it froze you would have ruptured the cell walls; with a higher level of the ani-freeze it would prevent that. Got me thinking about saving off some yeast, and giving it a go.


QldKev


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## Nevalicious

Its definitely worked several times for me now... 

I will go with the suggestion Bribie that you didn't have enough glycerine. I really dont measure it out as such, but rather, roughly fill the vial (20ml) 1/3rd full of glycerine, then top up with yeast slurry/smackpack etc. I've had no issues thus far getting one going again. 

As per your post a few days ago, I generally do the first step up on about 300ml of wort, no more...

Anywho, just my 2.43 cents

Nev


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## Wolfy

BribieG said:


> It is with deepest sorrow that I report that the brave and patriotic yeasties did not make it through Cryo.
> 
> I suspect my freezer just wasn't cold enough, well onto some slanting now and see if that works better - have the agar.


I'm very surprised that no yeast remained viable while it was frozen.
Rather than not being cold enough, my guess is the problem was with freeze-thaw cycles or something else.

I've been freezing yeast since last year - in small cultures, about 30% glycerin, and a method closer to what is outlined in the 'Yeast' book - while some (might even be strain-dependent) has suffered obvious losses, each of the dozen frozen samples still has some viable yeast.

Slanting yeast is easy, but it does get to be a-lot of time work and effort, so after reading the book, washing yeast and storing it under (distilled) water is what I'd like to try next for long term yeast storage.


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## felten

MHB said:


> removed the superannuant liquid (beer)


the pensioner liquid? :lol:


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## davo4772

Sounds like better have too much glycerine than too little.


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## Bribie G

I must admit I was reluctant to put in too much glycerine as I thought it might taint the beer - OK I will definitely try it again and up the G content.
I have _n_ packs of 1469 where _n_ is a large number and I'll use one of these as an inoculator. I really want to get the method nailed because even though you can resurrect a smack pack after a few months, waiting 2 years for the next release is a bit of an ask. Plus I'd like to have stock to help out other brewers as they have helped me out (Bribie goes out to car to get violin) 

Cheers guys, feel heaps better after your advice. :icon_cheers:


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## MHB

felten said:


> the pensioner liquid? :lol:




OK, ok - try Supernate God dam spell checker mutter mutter grumble 

MHB


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## Wolfy

BribieG said:


> I must admit I was reluctant to put in too much glycerine as I thought it might taint the beer - OK I will definitely try it again and up the G content.
> I have _n_ packs of 1469 where _n_ is a large number and I'll use one of these as an inoculator. I really want to get the method nailed because even though you can resurrect a smack pack after a few months, waiting 2 years for the next release is a bit of an ask. Plus I'd like to have stock to help out other brewers as they have helped me out (Bribie goes out to car to get violin)


By using smaller samples of yeast and being prepared to step-up a starter means you can use more glycerin without it being a problem in your beer. I've just finished testing the viability of my own yeast freezing experiments it's much more labor intensive and time consuming than your way, but by using small samples the viability is more easily. It just needs more work and then a starter to grow pitch-able quantities from the very small number of cells that are revived after being frozen.

I also think that yeast will 'keep' in their sterile packs much past the use-by date, but once again you may need to start with small starters or even streak the yeast out onto agar once very old packs are opened.


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## davo4772

Just froze 80mls of top cropped 1272 with 25mls of glycerine in a sterile 500ml glass bottle.

Not very scientific, about 30% glycerine. 25mls

Sitting in a small esky with another block of ice.

Has not frozen solid, still in liquid form.

Will leave it a month or two and see how it goes.


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## argon

argon said:


> Froze me some yeast last night :beerbang:
> 
> Tried it first with some good ol' US 05... i figure if things don't work out no loss. If it's all good i'll be freezing some precious 1469 i have in the fridge.
> 
> Couldn't have been easier... especially considering i used sample jars and syringes that are pre-sterilised in the packet.
> Just made up my solution of Glycerin and pre-boiled water, brought it to boil again in microwave, added to sample jars, added appropriate amount (35ml in my case), then took some top cropped washed yeast and syringed 10ml each into 5 jars.
> 
> I now have 50ml of washed, frozen top crop US05 ready for starting up whenever i want. I can see this yeast harvesting and storage thing getting out of hand and filling up my freezer with sample jars of murky brown liquid. :blink:
> :icon_cheers:




Success! :icon_cheers: Made a starter out of my frozen US05 yeast from April over the weekend. 

Friday, took the frozen sample out of the fridge and let it defrost/come up to ambient over a few hours, then pitched to 200mL and stepped up after 24hrs into 2L. Yesterday evening had a nice thick creamy head on it... even when on the stir plate. Late last night decided to crash chill it after around 36hrs on the stir plate and this morning have a very nice 9-10mm layer of light yeast on the bottom of the 2L bottle. I'll be pitching tomorrow night into 40L of 1059 wort.

Got another 5 x US05s, 3 x 1469s and 3 x 3522s in the freezer. I'll certainly be doing this from now on when wanting decent yeast storage.


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## Bribie G

I'll have another go at ressurecting some of those tubes - I may have shocked the last lot by not thawing over 24 hours. Poor planning resulted in my rushing the yeast.


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## davo4772

I froze 30mls of rinsed ESB 1968 yeastcake with around 12 mls of glycerine (40%) Just under 2 weeks ago.

It remained in liquid form. Just pitched into an ESB after stepping up with 200ml, 1L, 1.5L. Starter fired up like any othe starter I have done. 

Was only at minus temperatures for 2 weeks but the yeasties survived the freezing process no problem.

I also froze some 1272 with less glycerine, between 20 and 30%. Those samples have frozen solid so not sure how they wil go, no plans to make more APA's currently.


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## stillscottish

Just pitched a frozen 1007 into an Alt this morning. 
Let it thaw in the fridge then brought up to room temp and made a starter. Took 1 1/2 days to come to life. 

The first batch I froze failed. I suspect this was due to not enough yeast in the sample. I now let the yeast settle for a couple of days to make sure it's a good dense sample in the tube. 35 ml yeast topped up to 50 ml with glycerine.

Campbell


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## Midnight Brew

Bribie did you ever end up giving this another go?

Just gave this a crack with some Wy1318 London Ale III. My solution was a little less glycerin then whats recommended in this thread so hope all goes well. As it stands its frozen solid in my freezer. Have 3x vials with about 70billion cells in each.

I used this method: http://www.homebrewtalk.com/entries/freezing-yeast.html


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## Yob

he. he.. timely post.. doing this today myself 





15% solution ready for the yeast :super:


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## Bribie G

Best of luck guys, I didn't replicate the experiment. I do a limited range of styles nowadays and find that three or four yeasts keep me truckin along and I keep them in PET bottles of slurry for repitching within a month or so. :beerbang:


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## Midnight Brew

Wicked tubes Yob! Plan to get some more 1318 into tubes tomorrow night then in a few weeks some 3068 when I ferment the other half of my weizen and then depending if she can be saved and cultured up, the mighty Greenbelt. Will keep some backups in the fridge just in case things go wrong. Collecting and freezing yeast could be very interesting.


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## Yob

Bribie, the starter I made was from a slurry of Greenbelt Ive had in the fridge since September 17, Ive run a starter and plan on getting 80ml of the yeast, 20ml into each tube to freeze. (the rest to pitch)

Its as much an experiment as it is wishing to save particular generations of the yeast., Say get 3 from successive uses and freeze. We all know yeast tend to hit a sweet spot with usage and I intend on finding out what it is for Greenbelt. 

Id also like to build a comprehensive bank of the various yeast I use for APA, IPA, AIPA, etc.. Ive tried withslurry and it tends to get a bit long in the tooth and requires too much work to keep them all fresh.

I love these sorts of experiments anyway so win all round. Plus, it'd be good to have enough saved to be able to swap yeasts at Case swaps, a practice that seems to have died out in the last few years..

h34r:


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## Grainer

if you can freeze it quicker you will get better recovery as sending it to -20degC slowly causes shearing.. dry ice and ethanol/methanol is the way to go if you can get your hands on it.


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## Bribie G

My original experiment was in a fridge freezer, now I have a serious chest freezer I expect it would go better in that.


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## Midnight Brew

Grainer said:


> if you can freeze it quicker you will get better recovery as sending it to -20degC slowly causes shearing.. dry ice and ethanol/methanol is the way to go if you can get your hands on it.


Will give this a try today by just going straight into the freezer. With current samples I've whacked them in the fridge to cool down to 4c first. I thought it was best to cool to 4c first to build up some trehalose but then again that is a slightly different method where ascorbic acid is used with a 50/50 mix of glycerin/ypd medium. p199-200 Yeast book.


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## MartinOC

> Wicked tubes Yob!


Timely!! I'm trying to organize a bulk-buy of these (& similar tubes) right now if you want to join the party:

http://aussiehomebrewer.com/topic/77474-melbourne-sterile-sample-tubes-yeast-storage/


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## Yob

I made a little foam shielded container today and put my first ones in.. had a look before and they look to be sitting at about -10'c

The 50ml vials fit in perfectly so I shielded the walls on the inside and the base on the outside and applied a liberal coating of gaffa tape to it.

Looking forward to pulling one out at the end of the week and putting it into a starter.





ed: photo taken after they had been in the fridge overnight and most of the day and prior to going off to the deep freezer


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## Grainer

You will get better recovery rates from smaller vials as there will be less shearing of cells.. remember you have to freeze FAST!!


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## Yob

Yeah I know mate, but this is the equipment I have on hand, at the end of the day its essentially a no cost experiment (OK $10 for the Glycerine) so Ive not a lot to lose. 

If it remains viable and steps up its a winner, if it doesnt fire I will need to look into other methods or continue as I have been with slurry 

:icon_cheers:


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## MartinOC

Excellent stuff, Yob, but how does SWMBO feel about having one of her prized snap-lock food containers commandeered indefinitely for yeastie-beastie cryogenic experimentation, Dr Frankenfurter??


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## Grainer

Yobbo.. try freezing it in a slushy of ice water to get the temp down faster.. !!!.. flavour it with vodka and espresso and the wifey has a espresso martini afterwards.. Alcohol helps in dropping the temp during freezing .. LMAO


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## Yob

Those containers are mine... As is the freezer space..

Mostly she just rolled her eyes when I told her what I was up to .. She is a very understanding lady.. Bless her she needs to be


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## MartinOC

I can see her rushing to dial the number for those nice men in clean white coats when you start yelling "It's ALIVE! It's ALIVE!!" in coming months.....


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## Yob

Grainer said:


> Yobbo.. try freezing it in a slushy of ice water to get the temp down faster.. !!!.. flavour it with vodka and espresso and the wifey has a espresso martini afterwards.. Alcohol helps in dropping the temp during freezing .. LMAO


Wouldnt a salt ice water slurry do the same?


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## Yob

So I pulled one of of cryo after a week, left in the fridge for 2 days, and threw it on the stirplate this morning, 6-7 hours later I have this..




Looks viable to my eyes, will be interested to see it as the day unfolds.

:icon_cheers:


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## Yob

So Im going to call this a raging success, the photo below is at ~8 hrs, by 12 hrs it was trying to jump out of the flask,




at 24 hrs its all over, I turned the stir plate off and it's dropped back, significant change in the colour of the wort.




I could easily pitch this 500ml starter into a 2lt one and be ready to pitch to a brew, from start to finish, in 48 hours.

Overall, Im stoked, the yeast that the above is pictured was frozen solid for a week and to have it fire like that was astonishing fro a first attempt.

Cheers


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## Yob

soooo.. thinking snap freezing..

something like >THIS< ?

I get some sun spots due to my fine Scottish heritage that need to be frozen off every so often, got me thinking about application to yeast snap freezing.. current experiments suggest it isnt exactly needed but damn it'd be fun h34r:


----------



## Not For Horses

Just don't keep it near your deodorant...


----------



## Yob

Banking my second lot today, WY1217 and Ive got a little mini WLP001 to go next.. then the WLP High gravity and then WLP San Diago Super Yeast.. I think that'll about do me for starters ( -_- )

:super:


----------



## MartinOC

Yob said:


> I think that'll about do me for starters


I see what you did there...clever trousers!


----------



## Midnight Brew

Also going to call this a raging sucess.

My experience with Freezing Yeast.

I've frozen about 4 lots of yeast vials (1 being a fail) and have 3 different yeast in the freezer tubed up as we speak. What I've learnt through discussions with Grainer is the yeast need to be frozen quick, really fast. Otherwise the cells will crystalise and most likely kill them all. I did this once and a clear example too when held next to a vial frozen correctly.

2nd mistake I made was thawing out and putting into a starter. I put about 20 billion (5ml) yeast into a 1 litre starter and for what reason I thought that would work I have no idea. Now I give the vials a day in the fridge, 2 days in a 50ml starter thats shaken intermediatly then 500ml then what ever I need using calculators. YES it's a few starters at this stage because Im freezing amounts of around 7ml which is about 20-30 billion cells depending on factors. I will possibly get into freezing sizes of around 20ml yeast then it will just get a 1L starter which is just over the amount needed for 23L of 1.050 wort.

So far I've trialed 2 tubes of the same strain. One being the fail and the other being the sucess. Now more on the sucess.

Wy Greenbelt a bit of an unknown strain to most which was sourced from Wolfy about 2 years ago. Between Yob and myself its been kept going a few times. Yob gave me a vial which I thawed out, grew in a starter and used in an APA. From that APA I've harvested and rinsed the yeast and now frozen several vials. At this moment one of those vials is rocking away in a starter and is going into an AAA.

Got a batch of Bitter fermenting now with 1469 which will be top cropped and also frozen. This will take me up to 4 strains in the freezer at my disposal.

This is becoming its own hobby! So many options!


----------



## manticle

Inadvertently froze about 5 wyeast packs as my hop fridge thermostat has decided colder is better. I have since transferred to the freezer section of my ferment fridge which seems to stay cold but not frozen and I have brewed a few beers using thawed yeast (always using large active starters first).

So far beers have turned out well including some big (1070-1080) ones. One of them is a Czech pils - be interesting to see if I can get a good result.

Obviously I am not recommending freezing by throwing a whole pack in the freezer as is.


----------



## lael

Midnight, what method are you using to freeze quickly? I've seen the name greenbelt pop up a few times on here. What is it like? Want to swap for some Conan?


----------



## Yob

If MB doesn't, I do 

I'd not heard of conan till just now, 

Bit of a read up and it sounds pretty good. 

Greenbelt is pretty clean and drops like a brides knickers, I love it, haven't used recently as I've been expanding the library (001, 1217, 009 etc)


----------



## Midnight Brew

lael said:


> Midnight, what method are you using to freeze quickly? I've seen the name greenbelt pop up a few times on here. What is it like? Want to swap for some Conan?


Once all the tubes are setup and ready to go they go into an ice bath of water, ice and a bit of salt for a good 10 minutes. Then I place the tubes in a plastic container, fill it with some of the ice and then put into the freezer with a few ice bricks around it. The idea is to have the vials already at 0C or below when they go into the freezer.

WY Greenbelt is a great performer and I love 1272 but prefer greenbelt over it. It gives off this wonderful fruity character that complements American styles. Used it in a big stout about 6 months ago and chewed it from 1.067 to 1.009 in about 5-7 days. I have no doubt Yob's pushed it further then this haha.

I've got them now in 15ml vials (about 5-7ml yeast) and in a few weeks should have some more in 50ml vials (20ml yeast). Happy to send you some. I've got Wy Greenbelt, 1318, 3068 with 1469 on the way.


----------



## lael

I think I've got mine in 30ml vials. Not a ton in there. But have stepped it up successfully. What is 1318? Where was greenbelt from?

Yob, do you have anything else interesting? 

So greenbelt is not as clean as 1056 or US05? what temps is it good at? Fave styles to use it for?


----------



## Midnight Brew

1318 is Wy London Ale III. The greenbelt came from Wolfy (Melbourne AHB member) and if my memory serves correct he got it sent from Texas? It would have drifted alot through the years as we've kept it going between us. Still a great performer.

Here is some more on it: http://www.austinhomebrew.com/Beer/Wyeast/Wyeast-Greenbelt.html

I tend to use it around 20-22C and comes out beautifully. I've used it in American : Blondes, Pales, Ambers, Browns and Stouts. It was my house strain for about a year.


----------



## Yob

lael said:


> Yob, do you have anything else interesting?


San diago super yeast, 1217, 001,099 and the Greenbelt of course, still to get 1272 1056 and a couple of others...


----------



## lael

San Diego super yeast sounds special. Is that the anchor strain? What are 001 and 099?


----------



## pedleyr

lael said:


> San Diego super yeast sounds special. Is that the anchor strain? What are 001 and 099?


Super yeast (WLP090) is apparently a proprietary White Labs strain. It's a great yeast, it's resilient, fast and flocs better than the Chico strain. It is pretty neutral, probably a bit more towards malt focus than Chico. 

It chews through any sub 1.050 beer I give it in ~3 days. Definitely worth a try. 

I don't yet freeze my yeast but I have a starter of it going at the moment that I could chuck in a stubbie if you want to give it a try.


----------



## pedleyr

lael said:


> Edit:double post, apologies.


----------



## Yob

lael said:


> San Diego super yeast sounds special. Is that the anchor strain? What are 001 and 099?


All 3 are white labs yeasts, 099 is a super high gravity yeast, up to a stupid %25 abv


----------



## lael

pedleyr said:


> Super yeast (WLP090) is apparently a proprietary White Labs strain. It's a great yeast, it's resilient, fast and flocs better than the Chico strain. It is pretty neutral, probably a bit more towards malt focus than Chico.
> 
> It chews through any sub 1.050 beer I give it in ~3 days. Definitely worth a try.
> 
> I don't yet freeze my yeast but I have a starter of it going at the moment that I could chuck in a stubbie if you want to give it a try.


That would be amazing. Where are you located?


----------



## pedleyr

I'm in Melbourne, and for some reason thought you were too... Where are you? Postage may end up being more than a fresh vial, let's see.


----------



## MastersBrewery

Yob said:


> All 3 are white labs yeasts, 099 is a super high gravity yeast, up to a stupid %25 abv


How neutral is the 099? any other characteristics you've noticed not listed at white labs. the 25% is you stupid after consumption.


----------



## Yob

I cant say for sure, only what Ive read as it's still in the FV, and TBH, it isnt in there by itself so will be hard to tell, Batch one had T58 to start it off for the first 4 days and batch 2 had Greenbelt I think (or 1217) so both are a bit of a bastardised concoctions  (as is my way)

Mostly Im using it (099) to be able to handle the ABV these are going to get to h34r:

Future brews I'll do exclusively with it to gauge the profile but from what Ive read (well at least the bits that stuck in my head) it favours a malt profile so probably not as neutral as other Ale Yeasts


----------



## lael

Ah, I'm up in Sydney... seems like there are more brewers down in Melbourne...

Well, Let me check what I have in the fridge in vials. I can spin up some yeast from vials I've got and send a few vials down via express if people want to swap? I'm guessing a sistema box with foam and techni-ice/similar on the inside plus some vials should all fit in 1 500gr express post satchel?

I've got the DT yeast I cultured and Conan, and 3724, 3726 (saisons) I think. Maybe some others, but I'm not sure about viability... so would need to spin it up and re-vial it.


----------



## lael

Wow - what are you making? I've read about T58 for a pannepot clone and have some waiting in the freezer for that adventure


----------



## Yob

It's a Belgian(ish) monster brew, but a hop monster as well, more of a specialty mix of styles, kind of barlywine cross IIIPA with a hint of Belgian finished with NZ hops in the keg


----------



## wynnum1

anyone had a look at this http://suigenerisbrewing.blogspot.com.au/p/yeast-exchange.html


----------



## Yob

wynnum1 said:


> anyone had a look at this http://suigenerisbrewing.blogspot.com.au/p/yeast-exchange.html


dude must have some serious time on his hands


----------



## menoetes

Hey Bribie, thanks for the thread. It's been really helpful. Knowing that I can not only farm, wash and recycle yeasts but freeze them too has finally convinced me to enter the world of liquid yeasts.

I bought my first strain (wyeast 1272 American Ale II) and made up a starter that I worked up from from 500mls to 2 litres over a few days. Then I split it, half going towards my new brew and the other half getting frozen in two 50ml sterile (need I say 'unused'?) urine containers from work. It was fun, I'll admit it. 

My concern is that once the yeast settled/compacted down after some time in the fridge there was only what looked like a heaped teaspoons worth at the bottom of the liquid each of these containers.A layer about as thin a hb pencil lead. Does that sound about right or have I diluted out my yeast to much for future use?


----------



## MastersBrewery

put on a stir plate in a 200ml starter and work up from there, it'll be fine.


----------



## menoetes

Cheers Masters, I've been sweating about this for a while. Glad someone finally put my fears to rest


----------



## Yob

You may even want the first step to be less than that, 50ml then to 500ml, low gravity wort of about 1030, they have been in a pretty stressful environment and a bit of love won't go astray


----------



## Yob

Loving this bit of Kit, cant remember who I got it off a couple of years ago, but makes everything so much better to Sterilize.





The equipment Im using




Half Way




Complete ready for Freezing




Ive taken to giving it a shake up every half hour when freezing to stop it stratifying, seems to be a better freeze than some of the earlier ones.




Ive got frozen now,

Greenbelt
WY1217
WLP001

still to do

WLP099
SanDiago Super Yeast
WY1272


----------



## geneabovill

Interesting stuff.. Seems I'm gonna need lab supplies added somewhere in my signature. I figure it'll pay for itself what with the money I save from _not_ buying three smack packs to pitch because I can't be arsed making a starter. 

Along with more fridges, freezers and other assorted paraphernalia. 

Subscribed - and I'll be giving this a crack in the near future.


----------



## Yob

mate, if you are freezing yeast.. starters you will need 

Stir plate highly recommended.. look up the Digital Stirplate, cheap and the most quality piece of kit for this sort of thing.. cant praise their goods high enough :super: 

In fact HERE is the link.. no affiliation just a happy customer, they even have a cheap DIY kit available, but if I had my time again Id go the plug and play again... a thousand times h34r:


----------



## geneabovill

Oh, I've got all the gear, I just haven't really had a reason to. 

Now that bub #4 is almost due, I need to save some cash, so now it's a viable option. 

The digi stirplate is next on my list, because I'm not heaps happy with my ghetto stirrer.


----------



## Yob

so with a bit if further research and testing ive decided to go with a simplified approach.

%50 Glycerine solution means that the actual tube doesnt freeze at all hopefully meaning its a bit better on the yeast in the samples and it's a lot easier to manage the whole process.

was wondering if I now *NEED* on of these... I assume that it'd help separate the yeast in the tube allowing to pour off the glycerine/water?

not sure need is the right word but damn it'd make the shed look a lot funkier B)


----------



## Mardoo

Why do you need a milking machine in your shed Yob? :huh: Hey, that could be fun for the swap day by the look of it!

Everything I've found in the last two days indicates it's fine to make the starter with the glycerine included.

Oh, and Auroma sell bulk glycerine for $19.25/litre or $66.00 for 5 litres.


----------



## Eagleburger

I tried 1:1:1 glycerol:water :wyeast slurry and it is working so far after a week .I thawed a sample this morning and put it in dme solution and they are going great.I  am sure I can see them smiling. I have about 100 1.5ml samples in the freezer.


----------



## Eagleburger

phone didnt add pic of the happy little yeasties


----------



## Yob

I was creating a new bunch of Greenbelt %50 solutions as I only have 2 of the original freeze, most given away with good feedback,

it was some months ago so Im going to thaw one of them and give it a run, the initial step will be just decanting the Glycerine / Water off the top of the tube and topping up with wort, then on to a 500ml starter then the 2000ml starter over the next few days.

These have been frozen fro approx 4.5 months.


----------



## Midnight Brew

With your 50% solution then added to the yeast, would that would give a final solution of 25%? (50% yeast then 25% water and 25% glycerine)

If I remember this right (dont have my notes in front of me) frozen vial thawed in fridge, then 50ml, then 400ml then a 1.5L starter and it worked a charm. I think it was a 5 day ferment too.

My freezer is full of yeast and hops. I clearly dont brew IPA's.


----------



## Yob

Midnight Brew said:


> With your 50% solution then added to the yeast, would that would give a final solution of 25%? (50% yeast then 25% water and 25% glycerine


actually MBrew I think it's more %50 Glycerine, %25 Yeast and %25 Water, as can be seen in the pictures the Yeast slurry is filled to the 25ml mark, Glycerine then to the 50ml mark


----------



## MartinOC

Yob, just how "thick" is the slurry you have in those tubes above? It seems you're assuming that it's 50/50 water/yeast up to the 25ml mark, then you add 25ml of "straight" glycerine to the tube?


----------



## Yob

Roughly speaking, yep


----------



## tateg

Hey Yob
How's it going with the 50% Glycerine mix?, I have read somewhere else that raising the Glycerine % is the best option when using a frost free freezer.
Does the yeast have a chance to settle out before freezing ?
cheers


----------



## Yob

The solution doesn't freeze at %50 but yes will still settle slowly.

I haven't had to revive any ay the higher concentration yet but I fired up one of the %15 ones this week after being frozen for 4 months.. Fired without issue 

My freezer isn't frost free


----------



## lael

Sydney:
http://www.seoc.com.au/product.asp?pID=4017&cID=429 cheap food grade glycerine. These guys are cheapest by far. 

Noosaville, Qld: 
https://www.escentialsofaustralia.com/php/product_listing.php?gid=5 
Vegetable pharmaceutical grade is cheaper than above, actually...but have $8 surcharge under 100



Melbourne:
There's also n essentials 
https://www.n-essentials.com.au/mobile/product.aspx?ProductCode=RSGLYC&CartID=1

And http://www.melbournefooddepot.com/?rf=kw&kw=glycerine


----------



## Yob

Hay, good searching, that last link is a corker,

Comparison is 200ml = $8 at Coles


----------



## lael

Yeah, although the seoc is $8/kg 15 delivery in Sydney, and maybe for Melbourne too. I tried 2kg and it was still 15. So.. 31 for 2kg. Pretty awesome.


----------



## idzy

Or $29 for 5kg, sounds like a bulk buy opportunity.


----------



## Yob

Just to add to this thread, I stepped up some WY1217 last week and I pitched it on Sunday, brew is now sitting at 1010 so Im really happy with the continued results.

For a low cost, low effort method of capturing yeast, Im stoked with how this is turning out.

:super:

(ed I should add, been frozen solid in a %15 solution for 4 or 5 months)


----------



## kcurnow

Hi Yob, a quick question on sterilizing your syringe for transferring the right amounts of yeast and glycerine to the storage tubes. How do you sterilize them? Is it done in your pressure cooker or just with starsan?


----------



## Yob

I actually dont use it any more, I typically just pour from the starter flask to the storage tube and then glycerine right in on top of that.

I figure the simpler I can make the process the less cleaning I need to do.

but when I did, it was both the pressure cooker and then a bath of starsan.


----------



## tateg

hey Yob,
how many viable cells do u recon you get in each vial ?
are you still using %50 glycerin ?


----------



## Yob

I've not done cell counts or anything fancy like that mate, suffice to say, I've not had any issues stepping from thawed to 50ml then 500ml then 2lt, last ferment from what was my second freeze, (4-5 months old) finished in 4 days.

Still doing %50 but the above was a %15 solution, will pull out a %50 one this week and see how it compares


----------



## tateg

Yob said:


> I've not done cell counts or anything fancy like that mate, suffice to say, I've not had any issues stepping from thawed to 50ml then 500ml then 2lt, last ferment from what was my second freeze, (4-5 months old) finished in 4 days.
> 
> Still doing %50 but the above was a %15 solution, will pull out a %50 one this week and see how it compares


Thanks yob
I am just trying to work out how much i should step up before splitting, i might just try for 100 billion in each vial


----------



## lael

tateg said:


> Thanks yob
> I am just trying to work out how much i should step up before splitting, i might just try for 100 billion in each vial


What rule of thumb are you following to count? Or are you actually counting?


----------



## tateg

Planing on using a yeast calc to estimate the cell count before freezing, using virgin yeast pack


----------



## Yob

Yes, I put them in 1 vial...

Not cells no, I count the mls if that helps, each vial contains no less than 10ml yeast (some more)

I could, if I could be arsed, back track it to Mr malty for a rough cell count but I feckin hate maths...


----------



## kcurnow

If anyone is interested Aldi are selling 6lt stainless steel pressure cookers for $45 at the moment. Perfect for sterilising. 


Sent from my iPhone using Tapatalk


----------



## Eagleburger

Four days ago this was 1/2ml of yeast slurry stored at <0 degC. Now its 120ml of compacted yeast ready to pitch in the morning. 

I believe this huge growth is attributed to the dodgy stirrer used. It gets crazy hot.


----------



## lael

Wow. Nice. How many steps?


----------



## Eagleburger

Three steps

Centrifuged tube, Added yeast pellet (size of matchhead) to 10ml wort kept at ambient ~ 20degC for a day. Got impatient and taped tube to back of warm television. it smelt like beer in another day, so two days total.

Added the tube to 200ml wort and put on warm stirrer. Twelve hours later had beer. 

Added 2L wort. 24 hours later had beer so put in fridge for the day. I could have pitched last night when I took the photo except the wort had not yet cooled enough. Pitched this morning at 7.5degC and set the temp for 9.5degC


----------



## geneabovill

Any ideas where I can get a decent inoculation loop, or similar tool for this sort of business?

I'm in Newcastle, NSW.


----------



## Kingy

Most online mushroom growing supplies shop sell inoculation loops.


----------



## geneabovill

Cheers mate. I'll check them out. All the lab supply joints want a $100 minimum order...


----------



## Eagleburger

make one from some ss leader wire.


----------



## Yob

Why do you need one for freezing?


----------



## geneabovill

More for streaking/slanting, buy couldn't find the thread for that topic... Plus I figured The God of Yeast started this one, so would know.


----------



## Yob

Wolfys not here much these days


----------



## Yob

http://aussiehomebrewer.com/topic/47107-making-agar-slants-in-pictures/

^^^ Slants thread ^^^


----------



## idzy

How'd that 50% go Yob, I am about to chuck some Glycerine in. Should I hedge my bets and go 1 each way?


----------



## Yob

Just threw in a Greenbelt last night that had a %50 mix... It's now making a horrible mess all over the kitchen table..

Ok I guess


----------



## idzy

Delightful


----------



## Yob

I pitched 20ml into a 2lt starter, it's going bananas after 24hrs... 

Destined for my mandarina Bavaria cube


----------



## idzy

So I have done a mix of 20ml glycerine and 25-30ml yeast into 10 tubes.

Put them in the fridge last night. Gave them a shake this morning and placed them in an open container with ice laid over the top.

Will keep all posted on success.

The yeast is Wyeast 3787 - Trappist High Gravity Yeast


----------



## Yob

an open container? I put mine in a sealed container with an ice brick

If you want some 001, 002, 090, 099, 1318, 1217, 1728, Greenbelt, er... and some other ones... lemme know


----------



## mofox1

Yob said:


> an open container? I put mine in a sealed container with an ice brick
> 
> If you want some 001, 002, 090, 099, 1318, 1217, 1728, Greenbelt, er... and some other ones... lemme know


Guessing this is to help maintain constant temps?


----------



## Yob

Nope, it'll swing more not less


----------



## idzy

I figured the ice would help cool down and the open container would allow cold to get in quicker. I guess I will put the lid on later, that's what I was thinking anyways. Otherwise the ramp will be slower I thought.


----------



## mofox1

Yob said:


> Nope, it'll swing more not less


Ah, my bad for not reading enough of the posts. I thought this was in the freezer, not while still cooling down. 

Would this be a good thing to do (for long term storage) if you have frost free fridge/freezer? Thinking it might help reduce the effect of the warm cycle.


----------



## Yob

well.. bit delayed on answering this Mofox..

Yep, great for frost free environments, i considered (still am) loading up a container with water in the test tubes, filling the container so its a 'brick' and then pulling the tubes out.. sort of like an Ice Test Tube Rack if you catch my drift?

I pulled out some San Diego Super Yeast yesterday, made a 500ml starter, put it on the stirplate this morning and the flask is full of condensation this evening, been 'frozen' (~%50) for 3-4 months

:super:


----------



## mofox1

Yob said:


> well.. bit delayed on answering this Mofox..
> 
> Yep, great for frost free environments, i considered (still am) loading up a container with water in the test tubes, filling the container so its a 'brick' and then pulling the tubes out.. sort of like an Ice Test Tube Rack if you catch my drift?
> 
> I pulled out some San Diego Super Yeast yesterday, made a 500ml starter, put it on the stirplate this morning and the flask is full of condensation this evening, been 'frozen' (~%50) for 3-4 months
> 
> :super:


Ha. No worries... I've since caught the freeze bug anyway. Using a 3L ish polystyrene box in the freezer to counter the warm cycle. Have several samples of 4 strains frozen already, with others in the fridge.

I reckon it might be hard to get a single tube out of an ice brick unless you liberally lube it up beforehand... Just, err, make sure you do it in private. Some things are just too hard to explain.


----------



## Yob

I dunno mate, the outside of the vial is pretty smooth, I'll give it a bash and let you know.. I reckon the friction will be pretty minimal


----------



## Midnight Brew

This is a first generation top crop frozen back in January of WY1318. This was from a 50ml vial which had 20ml yeast estimated at 70 billion cells, frozen very fast with a 15% glycerin solution.

I took it out of the freezer and popped into the fridge overnight. Then put it into a 100ml starter which I shook and swirled everytime I walked past it for three days (I might need a smaller flask so I can stir plate it). Then it went straight into a 1.5L starter and this is where she's at after 24 hours.








This is from the second batch of yeast I froze. 10 months in the freezer and she's hungry for some porter.


----------



## Alex.Tas

This is a rally cool topic. Thanks Bribie for starting it and everyone else for keeping it going.




Midnight Brew said:


> This is a first generation top crop frozen back in January of WY1318. This was from a 50ml vial which had 20ml yeast estimated at 70 billion cells, frozen very fast with a 15% glycerin solution.


When calculating yeast population growth from your defrosted starters, what sort of viability do you use? 
I use http://www.yeastcalculator.com/ when building starters.

for example: 
I made up a few starters of 3068 and split the slurry so that I estimate that I had 68b cells. The "production" date of the yeast I use is when it hit final gravity in the starter, which was say 15/7/14. The calculator tells me I have 46% viability after storing it in the fridge and therefore only 32.3b viable cells remaining so I base my starters off that initial quatity of 32.3b.

In your case though, the time between the final gravity of the starter and when you are restarting your 70b was spent near-frozen. So what baseline/viability do you use for defrosted yeast?

Or is this a silly question because it's an inexact art when predicting yeast populations from starters without doing a proper cell count? 
Maybe i'm getting too hung up on pitching mathematically/theoretically the right quantity of yeast...


----------



## Midnight Brew

I have more or less winged it and assumed the yeast is only a little less viable then what it was when I cropped it. I presume this because once its cropped I wait a few days for it to floc down then add the glycerine solution. By this time I've lost some viability naturally with time and then I've frozen it presuming I've lost a little more. At -14C and kind of suspended in time, the viability change would be minimal.

I could be very wrong but damn that krausen looks so healthy.


----------



## Yob

Further reading..

>HERE< is the thread by Wolfy which is a slightly different method to the one discussed here..

there are other write ups not from this site like THIS one and through linked to THIS one..

also on HBT >HERE< and HERE

and HERE on BYO


----------



## Midnight Brew

Yob said:


> Further reading..
> 
> >HERE< is the thread by Wolfy which is a slightly different method to the one discussed here..
> 
> there are other write ups not from this site like THIS one and through linked to THIS one..
> 
> also on HBT >HERE< and HERE
> 
> and HERE on BYO


Yob's above post contains more then sufficient information in those links for anyone to try it out. Well worth the time to read and try it out in my opinion.


----------



## Alex.Tas

Thanks MB. i was thinking along the lines of 90% viability. Sounds like that is similar to what you have assumed.

Yob, thanks for the links some useful info there but only one of them actually answered my question. The BYO article http://www.homebrewtalk.com/freezing-yeast.html about a third of the way down. 7.5%-15% glycerine solution seems to yield upwards of 75% viability over "several" months. That's pretty promising. 

Wolfy's method involved selecting really low quantities before and after freezing and then building the population from that. While it likely worked for wolfy and is probably the best method, I would rather try and store and revive a larger population so i don't need to step up quite so many times.

The other ones gave a some good pictorial step by step guides and slight variations on the freezing technique. The http://www.ipass.net/mpdixon/Homebrew/Freezing%20Yeast.htm link gave the method in it's simplest form

So Yob and everyone else, what sort of viability do you assume from your revived samples?


----------



## Yob

I assume I can step it up even with a low viability.. a fellow member gave me a 100ml flask a month or so ago and I love the bleedin thing.. its so tiny but so perfect for an inital step up.. so

~10ml -25ml yeast to a 100ml (1030) starter (12hrs) > 500 starter (1030) starter (18hrs) > 2000ml starter (1040) (24hrs)

Plenty of time to make sure you are getting the activity behavior you are after from the starter, right smell etc..

Viability.. pretty bloody high I rekon.. some of these things go just as nuts as if they hadnt been frozen fro a year.. Ive just had a 1060 IPA drop to 1010 (WY1217) in just on 4 days I think.. I wouldnt expect better from the smack pack  (though I might from a third generation 1217.. awesome..)

By the by.. had a big yeast day.. SWMBO said I was obsessed...

1272 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
1450 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
1056 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
1217 (just to have a reset) smacked to be split and frozen tmoz.. thinking I need a generator backup.. means I have.. off the top of my head at least a dozen strains frozen off atm... some of them also have 3rd generation harvests as well.. h34r: :lol:

:beerbang:

ed: clarity


----------



## barneey

Have you guys not used yeasty balls?


----------



## Alex.Tas

Ah brilliant. So its close to full viability. 
Looks like the days between christmas and new year might involve some watching demolition man whilst freezing some yeast. Then I know that when they get unfrozen they will come out of hibernation fully charged like a badass and hopefully wearing yeast sized denim overalls, and then not pay income tax.


----------



## mofox1

Yob said:


> By the by.. had a big yeast day.. SWMBO said I was obsessed...
> 
> 1272 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
> 1450 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
> 1056 fresh pack split into 5 vials (25ml %50 Glycerine solution 25ml Yeast/Liquid)
> 1217 (just to have a reset) smacked to be split and frozen tmoz.. thinking I need a generator backup.. means I have.. off the top of my head at least a dozen strains frozen off atm... some of them also have 3rd generation harvests as well.. h34r: :lol:
> 
> :beerbang:


What is the benefit of smacking them before splitting?

I've got my first wyeast in the fridge in the moment (3724 - belgian saison) and thinking of splitting it before I do the starter for the batch... would like to know whether to smack or not.


----------



## Midnight Brew

You've been busy Yob! I had some 1450 that I was gonna trade you haha but you beat me to it.

You've got a very suited collection to your brewing regime and will have to steal/trade some from/with you soon. It is becoming another essential to the hobby of being able to ferment at your own leisure.


----------



## Yob

barneey said:


> Have you guys not used yeasty balls?


No, not seen them, they look quite small?


----------



## Yob

mofox1 said:


> What is the benefit of smacking them before splitting?I've got my first wyeast in the fridge in the moment (3724 - belgian saison) and thinking of splitting it before I do the starter for the batch... would like to know whether to smack or not.


Just to proof them really.. Considered collecting the little packs to add to the starters but in the end couldn't be arsed, don't think it makes much difference to be honest, maybe better not to but as I sit them in the fridge to floc first, meh


----------



## barneey

http://www.pro-lab.com/literature/microbank-sales-literature.pdf

Basically you select a colony to use from a petri dish, place colony into vial, shake, decant liquid & dispose - leave behind the balls in the vial, place vial in freezer.

Create starter from ball either direct & step up or via petri dish & step up.

I have some yeast given to me grown up from a yeasty ball a couple of years old + have in turn made some more balls up earlier this year to try an experiment out in early 2015. Useful if you want to keep a yeast or platinum strain.


----------



## Yob

barneey said:


> http://www.pro-lab.com/literature/microbank-sales-literature.pdf
> 
> Basically you select a colony to use from a petri dish, place colony into vial, shake, decant liquid & dispose - leave behind the balls in the vial, place vial in freezer.
> 
> Create starter from ball either direct & step up or via petri dish & step up.
> 
> I have some yeast given to me grown up from a yeasty ball a couple of years old + have in turn made some more balls up earlier this year to try an experiment out in early 2015. Useful if you want to keep a yeast or platinum strain.


Hay man, they look pretty trick.. but one of the reasons Ive gone down the road I have is to avoid slants and plating.. simply dont have the time for it. The current method allows for a vial to be taken out on a weekend and is generally ready to use by the next which is pretty good... though it must be said that its starting to take up a bit of room with 3 to 5 50ml vials of 13 or so strains now h34r:

do you take out a bead and use that so each vial has multiple uses? how many balls to a vial? are they autoclavable thereby being multiple use?

Very interesting none-the-less, might shoot them an email 

ed: oh no.. not another feckin bulk buy... :lol:


----------



## barneey

The 2ml vial contains 25 beads, so they have in theory 25 uses (as long as you are ultra clean in using them / taking a ball out).

The company might if your lucky send a free sample, or the UK homebrew shops have now started selling them / if you get really stuck and want to pay for international postage I might be able to send a vial to you.

I would however treat them as an exercise first before doing a group buy as they do require quite a bit of preparation.

To save money however you could always buy some autoclave 2ml vials and start splitting the beads. 

As I mentioned they are of more use for storage of a rare / rarely available yeast strain, unless you like making up plates / slants & starters


----------



## Yob

Ive sen the US mob on that PDF link an email to see what they come back with, asked all sorts of silly questions 

Where you located mate?


----------



## barneey

England uk


----------



## Yob

Ah yes, should have gathered. 

Do you plate them or just step them up from the balls?


----------



## barneey

Plate them, although another member on a different forum (UK based) has used a bead with a very small starter and had success, (just throw the bead in).

I`ll also quote someone on the thread that originally put me on these beads

"I have a "Hens Tooth" one that is 4 years old and growing after being streaked from a colony from the plate that said yeasty ball was placed upon and when I finally get round to it it's heading in Barneey's direction.

Yes they are possibly slower than a slant but then you are probably starting with fewer colonies. Just roll the ball round on a plate of decent medium and go from there."


----------



## Yob

I love the direction these sorts of threads evolve into :beerbang:

Thanks barneey


----------



## Yob

I bought myself a 55lt tub today to go in the deepest darkest corner of the deep freezer (my hop one so it's feckin cold) and was surprised to see it nearly filled. Admittedly I can save some space be getting a few more *better suited* clip lock containers, some of the existing packaging isnt space friendly.. buuuut..

Im close to having 20 types of yeast and nigh on 100 vials. :beerbang:

Also.. It seems that a %25 solution still freezes, not an issue really but I do like it if they stay fluid.

ed: *_*


----------



## Grainer

where do u get ur vials Yob?


----------



## Yob

MartinOC did a bb a little while back and there was a bb through Melbourne brewers as well, I can get the details for both if you need?


----------



## Grainer

all good i will try from work


----------



## Yob

Get me some too.. 




I have a problem..


----------



## Camo6

Ok, I'm gonna get organised soon and give this a go. Where does everyone source their glycerin from? The local chemist only has 100ml for $5. I do have a bottle down the shed we had for one of the dogs but not exactly sure of its suitability.
I thought I'd try pressure cooking the glycerin in the vials first to sterilise. Can I safely do this with the lids screwed tight?
Also, does anyone use ascorbic acid as mentioned in the yeast book? Sorry if its been mentioned already.


----------



## MartinOC

How much do you need?

http://www.ebay.com.au/sch/i.html?_from=R40&_trksid=p2047675.m570.l1311.R1.TR5.TRC2.A0.H0.XGlycerine&_nkw=glycerine&_sacat=0

Edit: The 50ml tubes you got from me in the BB are autoclavable. Leave the lids just lightly screwed on. Once they're cool, you can screw-down the lids without fear of contamination of the contents.


----------



## Yob

Currently, I get mine from Coles, 200ml for ~$8 

Was talking to mardoo about this the other day and it's a possible bulk buy for us locally. I know mardoo has looked into it and has some options


----------



## Camo6

Cheers Martin. Looks like those prices are pretty comparable to the chemists. I might have to try the vets!

Those vials are half the reason I'm planning this. I'd forgotten about them until recently. A bit like all the other brewing related stuff I needed but yet to use!


----------



## Camo6

Admit Yob, you've got a problem!


----------



## Yob

I do indeed, mostly related to remaining freezer space and the amount of yeast varieties I'm still to bank


----------



## mofox1

Yob said:


> Get me some too..
> 
> 
> 
> 
> I have a problem..


Wait, wha? No bulk buy? Admit it Yob, you're slipping. h34r: 

I got 25 off Idzy a while back thinking there is *no way* I'd ever need more (contrary to the advice of Mr O'C & others IIRC). Split a saison last night leaving me about 4 empty's... now I think I'd better get more before I run out!

Next purchase will be for about 50. Or 100.

Dammit.


----------



## menoetes

Yob said:


> I do indeed, mostly related to remaining freezer space and the amount of yeast varieties I'm still to bank


I know how you feel Yob, when SWMBO decided we needed a chest freezer for under the house all I could think was 'Thank God, more room for yeast and hops'

I use the same 30% glycerine solution as in the tutorial at the start of this thread and clean (unused) 70ml urine specimen jars from my work when freezing. I too am growing a yeast bank and currently have;

3 x 1272 - American Ale II
3 x 1318 - London Ale III
3 x 2308 - Munich Lager
2 x 3724 - Belgian Saison
2 x Coopers Commercial Strain Yeast (recultured from bottles)

Going to add 1450 - Denny's Favorite 50 to that collection this week and working on finding a decent wheat beer strain to add to the collection too. Anyone know if Ferals White Witbier is primed with a wheat or lager yeast?

I'm open to suggestions on a good wheat yeast strain...


----------



## Mardoo

Best price I've found, we can get it for $13.20/litre if we buy in 5 litre lots, $12.10/litre if we buy in 20 litre lots. Provide your own bottles. I'd say minimum of 1 liter. Shall I seek the BB blessing?


----------



## kcurnow

I would be up for a litre


----------



## Camo6

I'd grab a litre too.


----------



## tateg

I would go one as well


----------



## MartinOC

At the risk of incurring the wrath of the minister for finance, war & saying "NO!", I'll go a litre too.


----------



## Yob

I'd say do it mate, I'll get at least 1.. Maybe 2


----------



## mofox1

Lol... just bought 2 x 250ml at Coles on the weekend. But whatever. Yes.

1L will do for what, 50 freezes? (@ 40%) So maybe two as well... gah. Expression of interest, err, expressed.


----------



## Mardoo

Hey y'all, I've put up the glycerine bulk buy thread here. If you indicated interest in this thread I put you down for the amount you said you'd like. Maybe visit the thread just to make sure I haven't portrayed your desires in a politically incorrect way, or something. If you don't actually want to participate just take yourself down or send me a nasty PM and I'll do it.


----------



## Midnight Brew

Added these to the bank today:

WLP028 Edinburgh Scottish Ale
WLP041 Pacific Ale

Next on the list:
WLP013 London Ale
WLP004 Irish Ale

That will take me to 8 strains. I feel a wrath of session ales coming on this summer.


----------



## mofox1

Midnight Brew said:


> Added these to the bank today:
> 
> WLP028 Edinburgh Scottish Ale
> WLP041 Pacific Ale
> 
> Next on the list:
> WLP013 London Ale
> WLP004 Irish Ale
> 
> That will take me to 8 strains. I feel a wrath of session ales coming on this summer./quote]
> 
> What did you use the pacific for? Have a vial I need to use or freeze soon, tentatively slated to go into a mild amber APA.... If I chicken out on the pacific it will just get a neutral strain.


----------



## Yob

In the interests of this thread, and the new bloody addiction of collecting every yeast on the planet... I've ordered 2 x 3lt schott flasks from digital home brew, I've now got a V3 and a V2 thanks to their awesome customer service.. Which brings the flask collection to

1 x 100ml
2 x 500ml
2 x 1000ml
3 x 2000ml
2 x 3000ml

Ok.. I might have a problem 

Cry havoc on the left and Greenbelt on the right, the Greenbelt was one of the first ones I froze


----------



## Mardoo

Pacific goes well in an American Brown.


----------



## niels

I've been lurking over this thread for some time and I've gathered most of the materials needed. The only thing missing is an inset for my pressure cooker (an old one from my mom) so that nothing is standing on the bottom surface.

Since I have 2 Wyeast smackpacks in the fridge that I will not be able to use very soon I was thinking of freezing them. I do have some last questions, though:

Does anyone knows how much yeast (in ml) is in a smackpack? It says "125ml", but is that including the nutrients?
Should I smack the packs and let them swell before freezing? Or only freeze the yeast that is in there? (I want to avoid making a starter if possible...)
Am I correct that you should make a mix of yeast, glycerine and water or beer?
What is the "optimal" ratio of yeast, glycerine and water/beer?
Voila... If I know the answers on these questions I'm ready for a frozen yeast bank!

Cheers!
- Niels


----------



## Midnight Brew

mofox1 said:


> What did you use the pacific for? Have a vial I need to use or freeze soon, tentatively slated to go into a mild amber APA.... If I chicken out on the pacific it will just get a neutral strain.


I havnt used it as of yet but initially bought it for use in brown ales and might even go in a blonde ale, and at some point no doubt it will go in a red beer (thats what the 004 and 028 is for also).


----------



## Midnight Brew

niels said:


> I've been lurking over this thread for some time and I've gathered most of the materials needed. The only thing missing is an inset for my pressure cooker (an old one from my mom) so that nothing is standing on the bottom surface.
> 
> Since I have 2 Wyeast smackpacks in the fridge that I will not be able to use very soon I was thinking of freezing them. I do have some last questions, though:
> 
> Does anyone knows how much yeast (in ml) is in a smackpack? It says "125ml", but is that including the nutrients?
> Should I smack the packs and let them swell before freezing? Or only freeze the yeast that is in there? (I want to avoid making a starter if possible...)
> Am I correct that you should make a mix of yeast, glycerine and water or beer?
> What is the "optimal" ratio of yeast, glycerine and water/beer?
> Voila... If I know the answers on these questions I'm ready for a frozen yeast bank!
> 
> Cheers!
> - Niels


1. I could be wrong but I think it is 25ml of yeast at 4 billion cells per ml = 100 billion cells.

2. I dont think it matters but generally if smacked it gives you a good indication that the yeast is alive and well so why not.

3. You want a mix of water and yeast (water + yeast = volume) then add onto that the percentage of glycerine you will be using.
Eg 40ml water + 40ml yeast = 80ml + 15% glycerine (12ml) = 92ml.

4. Depends on the resources you look at there are different answers. Plenty of brewers use 10-15% with success and plenty use up to about 50% with success. I originally read that over 15% can be detrimental to yeast and Yob has well and truely proved that to not be the case. Read up on the resources linked in this thread and decide for yourself :icon_cheers:


----------



## niels

Thanks for the quick reply.

Assuming 25ml of yeast in a pack I could go with the following: 25ml yeast + 25ml demi water + 10ml glycerine (20%) = 60ml
Having 20ml vials I can pour this in 4 vials at 15ml a vial...

Sounds like a solid plan, doesn't it? 

- Niels


----------



## Yob

From a smacked pack, I got 5 vials of 25ml which ended up about 5ml yeast in each.

To this I added a %50 solution of glycerine to give me a %25 final solution. At %25 they have still frozen solid, but that seems not to matter. 

Have a good read through the while thread mate, there is plenty of info on different ratios etc.

You will need to make starters.


----------



## niels

Yob said:


> From a smacked pack, I got 5 vials of 25ml which ended up about 5ml yeast in each.
> 
> To this I added a %50 solution of glycerine to give me a %25 final solution. At %25 they have still frozen solid, but that seems not to matter.
> 
> Have a good read through the while thread mate, there is plenty of info on different ratios etc.
> 
> You will need to make starters.


Sorry for my ignorance, but I'm not a native English speaker and sometimes I misinterpret basic things.

If I understand you correctly you have smacked the pack which resulted in 125ml of slurry. Divided over 5 vials gives you 25ml/vial with approximately 5ml of yeast per vial.

On this you added 25ml of glycerine mixture (50% glycerine and 50% water) per vial. This totals to 50ml per vial constisting of 25ml yeast slurry, 12.5ml glycerine and 12.5ml water.

Correct?

Regarding my starter statement: Of course I will have to create starters from the frozen vials, but I meant that I didn't want to make a starter of the smackpack and harvesting that yeast for frozen storage.

I started re-reading the thread and making notes, but I'm always nervous for my first time  So I wanted some confirmation and avoid flushing two healthy smackpacks down the toilet.

- Niels


----------



## mofox1

Nutrient pouch is around the 25ml mark. Chose not to smack, and got 3 50ml vials to around 2/3 full prior to adding glycerine.


----------



## Yob

That's all correct neils, spot on. 

I got some yeast this week cheaply that are just out of date, so I'm making starters for them to revive them and ensure they are at maximum health when frozen.


----------



## MartinOC

Yob, how big are you doing those "reviver" starters & how/do you allow for extra cell-counts in the resulting slurry?

Or, are you making a starter, decant/wash to ensure you're getting live yeast to freeze & discard obviously dead cells?

Or RDWHAH??


----------



## Yob

Technically I probably should rinse, you do get the opportunity from the starter flask, but to date I've not been overly concerned.


----------



## niels

mofox1 said:


> Nutrient pouch is around the 25ml mark. Chose not to smack, and got 3 50ml vials to around 2/3 full prior to adding glycerine.


Wow, this means it contains about 100ml of yeast and only 25ml of nutrients.

If so I can split a pack over a lot more vials.

- Niels


----------



## Yob

MartinOC said:


> Yob, how big are you doing those "reviver" starters & how/do you allow for extra cell-counts in the resulting slurry?
> 
> Or, are you making a starter, decant/wash to ensure you're getting live yeast to freeze & discard obviously dead cells?
> 
> Or RDWHAH??


From a vial or pack that isn't too far out of date, I go into a 500ml flask 10 ~1030 wort for 12-24hrs and then into a 2lt flask.



niels said:


> Wow, this means it contains about 100ml of yeast and only 25ml of nutrients.
> 
> If so I can split a pack over a lot more vials.
> 
> - Niels


You'll find that there is ~125ml total volume (inclusive of the pouch), with about ~25-30 ml yeast solids in that


----------



## mofox1

niels said:


> Wow, this means it contains about 100ml of yeast and only 25ml of nutrients.
> 
> If so I can split a pack over a lot more vials.
> 
> - Niels


^100ml of yeast suspension (incl water).

I wouldn't have wanted to split it over more than the 3 vials I did, it was looking pretty thin and after it had compacted down it looks like there's only about 5ml or so of solids in each.

Then again, maybe I got the runt of the batch...?


----------



## mofox1

Slightly OT, but all the interested folk are probably here... I'm thinking of buying some more 50ml tubes from proscitech.com. Including delivery, it's around 63c ea if I get 100, or 54c ea for 200 etc.

I would have preferred 25 or 30ml tubes, as the 50ml ones seem HUGE... but can't seem to find them anywhere. So 50ml it is.

Any interest in a bulk buy? Or are there better deals to be had elsewhere?


----------



## MartinOC

What you're looking for from Proscitech is the LS22-30 or 30L (labelled). Scroll down a bit.

I got some of those, but most people went for the LCA03 (50ml) in the last BB I organised through Proscitech. Bags of 50 for about $22??

WHATEVER you do, DO NOT got for anything made from PET & expect it to survive autoclaving (or even hot water). Chemical sanitation ONLY for those babies!!

I subsequently found a place local to me in Melb. that had the exact same things cheaper & I could collect, so no postage. http://www.fusionscientific.com.au/product01.html

Maybe check them out & ask for prices?

Alternatively, there's heaps of stuff on Evilbay, but check you're getting HDPE (ie. Autoclavable), which means they can be re-used & sterilised in a pressure cooker.

Freestanding tubes will make life easier for you, unless you buy a stand to work with them (once they're in the freezer, it doesn't matter).


----------



## mofox1

Cheers, missed those 30ml ones. Have already contacted fusion... Waiting.
Wasn't sure I wanted to go eBay for these, mostly because I want to have at least some assurance that they will be sterile, and made of the material they are advertised as!


----------



## Mardoo

Hey Martin, maybe you could pop those up on Yob's Bulk Buy Suppliers thread?


----------



## MartinOC

He's already beaten you to the punch by a couple of hours 

I'll get to it later this arvo.


----------



## niels

mofox1 said:


> ^100ml of yeast suspension (incl water).
> 
> I wouldn't have wanted to split it over more than the 3 vials I did, it was looking pretty thin and after it had compacted down it looks like there's only about 5ml or so of solids in each.
> 
> Then again, maybe I got the runt of the batch...?


Ok, it looks like I might run into troubles if I use my 20ml test tubes.

If I would use the yeast suspension straight from the pack and add glycerine to it I would end up with e.g. 100ml yeast suspension + 30ml glycerine = 130ml solution.

Dividing this over 20ml test tubes I will end up with 7 filled tubes but only 3.5ml of yeast per tube.

Do you think it is doable to dump this 20ml in e.g. a 30ml 1.030 starter as the first step for a starter?

- Niels


----------



## MartinOC

Absolutely, Niels!!

In theory, you can start anything from a single cell, so it's just a matter of stepping-up your starters until you have what you need. A stir-plate will make this easier.


----------



## mofox1

MartinOC said:


> I subsequently found a place local to me in Melb. that had the exact same things cheaper & I could collect, so no postage. http://www.fusionscientific.com.au/product01.html
> 
> Maybe check them out & ask for prices?


Prices for those guys are $7.50 for 50 x 50ml self standing, and $9.50 for the 50 x 30ml self standing... the 50ml has a screw on lid, but the 30ml is a "push" on lid (not too keen on that for some reason).

Postage $13 for up to 10 packs of 50.

I've emailed the guy for one pack of the 50ml... Still got time to add more to the order, so PM me if you:

Are in the eastern/south eastern 'burbs of Melbourne, or
Like a long drive for few dollars saving, or,
Can't resist the opportunity to see me in my natural environment


----------



## Yob

Make sure they can be autoclaved, and also have legible markings on the side.. If those are both winners, I'll take 50


----------



## MartinOC

I can feel a bulk-buy coming on...

I can feel a bulk-buy coming on...

Got the taste for it,

Just can't wait for it.......etc....

If you want to organise, I can collect, since this bloke is local to my folks' place (ex-local to me).


----------



## Yob

Yob said:


> he. he.. timely post.. doing this today myself
> 
> 
> 
> 
> 
> tubes.jpg
> 
> 15% solution ready for the yeast :super:


Late last week I thought I might take out one of my oldest, a Greenbelt now approaching 12 months from the look of it.. it was a few years old even then  ... anywhoo, it went into a 100ml flask for ~24 hours > 500ml > 36 hours and is now in a 2000ml flask, within hours of hitting the 2L flask it was showing foaming so overall quite pleased with the performance thus far. This is due to be pitched to a 1.060 IPA on the weekend, 

Ive got the case swap dark ale just finishing off with 1272 and its performed admirably, though it had been frozen for significantly less time I saw little difference in the starter activity times which Im quite chuffed about in terms of long term viability.

... FYI they dont ship international so if you plan on getting some you will need to sort out an alternative method than directly through the store


----------



## kcurnow

I have a question for all the yeasty experts here. I have an old (sep 12) packet of 3864 Canadian Belgian yeast that I want to revive so I can freeze it for extended storage. I have estimated viability of between 1-5% so was thinking of putting it through a couple of starters to build up the healthy yeast again. The question I have is the size starter to start with. If I go by the yeast book I should be starting with a 15ml starter to keep the inoculation rate around 67 million per mil (based on 1% viability leaving 1 billion yeast cells in the pack). That seems pretty small considering the packet will have 125ml in it however I guess most of that will now be dead yeast cells water and nutrients and I will need to rinse off the dead yeast once I have run the yeast through a few starters. Thoughts or comments on this approach? And recommendations on starter size? Cheers Karl


----------



## Yob

Probably a good idea to rinse as you say, once you have your rinsed volume, you will have a better idea of the starter size, If you put your rinsed yeast into a 50ml tube you'll get the amount of yeast measured, I've sometimes run the first starter in the tube, 1020 wort for older yeast, then onto 500ml (1030) then 2lt (1040)

Are you going to smack the pack? You might get a good idea of viability from that and could maybe go into a 1lt starter and rinse from that?


----------



## kcurnow

Yep pack has been smacked, waiting to see what will happen now. I might just put it into a 100ml starter and then rinse the result of that then go from there.


----------



## Midnight Brew

Hey Karl,

It is one of those situations where you dont have anything to lose except a bit of malt extract so I would go for it if you have the time. I think your calculations seem right if there is anything left in that pack and it could work. So on a stir plate steps would be as follows:

15ml
50ml
250ml 
1000ml by here you'll know if she is alive and if the calculators are correct you'll have close to 190b cells.

I dont think it would be worth the effort crashing and decanting each starter but then again you might have to depending on the size of flask and being there will be that initial dead yeast from the smack pack in there. It might be easier to have an initial flask and just keep feeding those increments to it every 24 hours.


----------



## kcurnow

What is the ratio of DME to water to give approx a 1020 wort as I normally make my starters around 1040 using the 100g/ litre ratio.


----------



## Midnight Brew

Half, 50g/litre is about 1020.


----------



## danestead

Just reviving some 1217 that I froze 2 months ago. Fingers crossed as this is the first one ive attempted to revive.


----------



## kcurnow

Well i have pitched it into a 15ml starter yesterday and will add another 50ml today.


----------



## Midnight Brew

I've got the Greenbelt up and going again courtesy of Yob's efforts getting some pure stock sent over from Texas. Pack is swelling and starter is ready and patiently waiting for a little more swelling. :icon_cheers:


----------



## Yob

I left mine overnight and pitched this morning onto 3lt starter, spinning happily away now, stoked to have pure samples again.


----------



## danestead

danestead said:


> Just reviving some 1217 that I froze 2 months ago. Fingers crossed as this is the first one ive attempted to revive.


Into the fridge for 12hrs then into a 200mL 1.020 starter then 500mL 1.040 and finally 1.5L 1.040 and all is going well.

Mixture on this batch was 50% glygerine/50% water solution then added in equal portions to the yeast therefore the final solution was 25% glycerine/25% water/50% yeast.


----------



## Yob

This is the ratio I've settled on as well. 

Freezing is cool


----------



## Midnight Brew

Yob said:


> Freezing is cool


Woah! He said it! He said it!


----------



## kcurnow

Well my 3864 freeze attempt hasn't gone too well. I am up to the 500ml starter and there is still no sign of any action from the old yeast. I have stuck a plastic bag over the top of the flask to see if it will inflate at all and give a hint of any fermentation activity. If not I will vomit and see if I can find some 3864 that isn't as old.


----------



## kcurnow

I have prepped a few vials with 50/50 glycerin and water (to the 25ml mark, to give me a final ratio of 25% water, 25% glycerin and 50% yeast solution once the yeast is added) and then pressure cooked them for 15mins to sterilise them. It was easy and now just need to add yeast. I am using WLP500 Trappist ale in the case swap brew so will harvest and freeze this.


----------



## Yob

I did the same with some wort about a month ago, just to see how 'sterile' a tube could stay, the wort vial has been on the kitchen window sill ever since and no sign of contamination. I'm quite tempted to do as you have done mate and pre prep the vials so all I have to do is add the yeast.


----------



## Camo6

Quick question fellow freezers. Added another 5 vials to the chesty yesterday and noticed 1 vial from my first frozen batch, the one against the wall of the freezer, was not frozen. Figured the freezer, which is on the decking, had warmed so I turned it up and moved the vials to the bottom surrounded by frozen bottles of water. Today the vial was still not frozen but the others were. What's up with that? The vial may have gotten an extra 5ml of glycerine to the others. Could that have made the difference. Fwiw I'm going with Yob ratios as posted above. Will check when this weather cools down to see if it's still fluid.


----------



## kcurnow

Camo6 said:


> Quick question fellow freezers. Added another 5 vials to the chesty yesterday and noticed 1 vial from my first frozen batch, the one against the wall of the freezer, was not frozen. Figured the freezer, which is on the decking, had warmed so I turned it up and moved the vials to the bottom surrounded by frozen bottles of water. Today the vial was still not frozen but the others were. What's up with that? The vial may have gotten an extra 5ml of glycerine to the others. Could that have made the difference. Fwiw I'm going with Yob ratios as posted above. Will check when this weather cools down to see if it's still fluid.


Hi Camo, I would suggest that the extra 5ml of glycerin will be the difference as in some of the earlier posts with higher %s of glycerin the mix wasn't freezing.


----------



## kcurnow

Yob said:


> I did the same with some wort about a month ago, just to see how 'sterile' a tube could stay, the wort vial has been on the kitchen window sill ever since and no sign of contamination. I'm quite tempted to do as you have done mate and pre prep the vials so all I have to do is add the yeast.
> 
> 
> 
> 
> 
> DSC_0063.JPG


Hi Yob, yep in one of the other yeast books "first steps in yeast culture - rajotte" he gives steps for prepping sterile wort in jars and sterile slants in exactly the same way. Then they can be stored at room temp. Just need to leave the caps loose during the steaming and then tighten once out of the pressure cooker and still hot.


----------



## Yob

Nice one, exactly what I did, sort of just wanted to 'proof' that it was as good a practice as you can get at home, kind of has in my mind, if anything was going to go bad in that wort, would have by now, especially being in partial sun.


----------



## kcurnow

Yob said:


> Nice one, exactly what I did, sort of just wanted to 'proof' that it was as good a practice as you can get at home, kind of has in my mind, if anything was going to go bad in that wort, would have by now, especially being in partial sun.


Yep and you need 15 minutes of wet sterilization under pressure. In the pressure cooker the steam should be at 121c at the highest pressure setting. It is the pressure bringing the temp up beyond the 100c boil mark that give the sterilization that boiling alone won't do.


----------



## Camo6

Brewnut said:


> Hi Camo, I would suggest that the extra 5ml of glycerin will be the difference as in some of the earlier posts with higher %s of glycerin the mix wasn't freezing.


Cheers brewnut, I was hoping that was the problem.

I've got jars of sterile wort in passata jars that are six months old and kept at room temp. If those crummy jars can hold a seal I'll have no worries about my vials. My $50 Aldi pressure cooker is one of my most useful brew tools.


----------



## MartinOC

Brewnut said:


> Hi Yob, yep in one of the other yeast books "first steps in yeast culture - rajotte" he gives steps for prepping sterile wort in jars and sterile slants in exactly the same way. Then they can be stored at room temp. Just need to leave the caps loose during the steaming and then tighten once out of the pressure cooker and still hot.


Ah, my first reading of yeast culturing techniques was Rajotte. Loaned the book to "someone" & never got it back :angry: .

I created agar "slants" in small McCartney bottles & pressure-cooked them, then screwed-down the caps once cooled. 

For those concerned about potential long-term storage of wort after pressure-cooking, relax. Those McCartney bottles were kept out of the fridge (in a shopping bag, shoved in a box) in my shed for 10+(?) years & showed absolutely NO signs of contamination whatsoever. I chucked them out when I purged for the move to Kinglake.


----------



## mofox1

Camo6 said:


> Quick question fellow freezers. Added another 5 vials to the chesty yesterday and noticed 1 vial from my first frozen batch, the one against the wall of the freezer, was not frozen. Figured the freezer, which is on the decking, had warmed so I turned it up and moved the vials to the bottom surrounded by frozen bottles of water. Today the vial was still not frozen but the others were. What's up with that? The vial may have gotten an extra 5ml of glycerine to the others. Could that have made the difference. Fwiw I'm going with Yob ratios as posted above. Will check when this weather cools down to see if it's still fluid.


Hey chief, I put mine in a small polystyrene box (from Coles, maybe 3 or 5 litres?) to maintain temps. Also an ice pack. I probably use a touch more glc as mine are mostly all liquid.... still yet to actually warm one up and revive.


----------



## Yob

Cam, a few% can make all the difference, %25 freezes, %30 does not as I recall (having played with mixture a bit) I wouldn't stress, at over %15 you're good to go.. I have one vial of 002 thatfroze among its compatriots that did not.. That worries me..


----------



## Camo6

Good news everyone! Just what I wanted to hear.

I've slowly filled the bottom of the chesty with frozen bottles of water as it's 3/4's empty most of the time, so this makes for a good nest for my vials. I'll look into making a decent storage container when I've done a few more packs. Cheers


----------



## Yob

Made a 3lt starter from my new greenbelt import, crashed it, made 3 vials and poured the rest back into a 2lt starter 

2 hours later


----------



## mofox1

Damn. I just turfed 5 vials from a washed us05 due to a suspected infection from the parent batch... So far I'm down five with none to show for it.

Serves me right for trying to save dosh on packet yeast.


----------



## Yob

You didn't turf the vials though right?


----------



## mofox1

Yob said:


> You didn't turf the vials though right?


At 15c ea, why not? h34r:


Nah, vials are safe and sound. Boiled the hell out of em too.


----------



## mxd

Hi Guys

I've read this thread (multiple times) and plan on doing my first attempt at freezing tonight.

I have 50 ml vials and plan on 25 ml yeast stuff from smack pack and 25 ml glycerin. (i've seen various combos in the thread this seems simple and easy for me.

Place it in the fridge for a day, then freeze AQAP (iced water etc.. then freezer with ice blocks around to help keep temp constant.

My Q's

I have seen it mentioned to boil the glycerin in the microwave, do you just put it in a cup and get it boiling, then let it cool and off you go.

1 of the packs I have I want for a brew I want to put down, so I thought I would freeze 4 vials then step up the 25 to 250 to 2000.

When I want to use a frozen I thought the 25 to 50 to 300 to 2000

When it's time to re-culture to restock do you use the slurry of a brew or do a 25 to 50 to 200 then to 6 or so vials ?


How does that sound ?


----------



## Yob

Hay matt, splitting from the pack, 25ml will yield about 5ml yeast solids in 5 vials, I step these up as follows, 

100ml > 500ml > 2000ml 

I can and do also freeze some off the slurry (rinsed of course) if the ferment was excellent so I'll end up with vials of different generations from original to about the third.

Ed: photo added 




This is some 099 I took out of the freezer yesterday, 100ml overnight and into the 500ml this morning, seeing some action on it now, will go onto the 2000ml tmoz


----------



## Yob

Side note, I've reduced my glycerine to %25 

12.5 ml glycerine 
12.5 ml water

Autoclave 

Add 25ml yeast from the pack.


----------



## kcurnow

Ok I have frozen my second batch of vials now. Included are OYL-500 Saisonsteins Monster, 3655- Schelde and 1217 - west coast IPA. I'm looking forward to using the 1217 in the Christmas Case swap cube I have sitting in the brewery.


----------



## Yob

thinking of doing this




when pulling out the tub to retrieve a vial, I worry about the temp swings, I naturally do so as quick as possible but part of the issue is that the first 50 or 60 vials I froze are written on the side of the vial not the top so identifying the vial I want can take a little time.

I hope this, in part, is a solution to a few of the issues, I can open the tub and lift out a "shelf" of vials with little to no temp swing, if I do get a power out, it'll last for days (brand new freezer so not worried about it carking it) 

I should be able to get 2 shelves in the tub which should hold about 80 vials each shelf.

should do me... :beerbang:


----------



## Alex.Tas

Yob said:


> I should be able to get 2 shelves in the tub which should hold about 80 vials each shelf.


Dude, you have a problem.





And I think it is contagious.


----------



## Yob

sooo, I went to do the 'ice shelf' thing the other day, got about half way the first time, knocked one over and the lot went down like domino's.. curses..

did it again, got all the way to the very last one and effing did it again... curses the greater.. turns out each layer will hold about 95 vials if I can ever get them in without knocking them all over..

anyway, I wanted to do an inventory of what Ive got frozen off already and get it on a spreadsheet so I can maintain it, though I'd show how easy it is to amass a decent library.

WY1056 3
GreenBelt (New) 7
Greenbelt (Old) 2
Greenbelt (3rd Gen) 4
WLP830 4 Wy1318 4
WY1318 8
WLP Burton 4
WY1217 8
WLP002 4
WLP041 1
WY1275 5
WLP013 1
WLP028 4
WLP545 4
WLP575 4
WY1272 3
WLP CRY HAVOC 6
WY1450 5
WLP001 3
WLP099 3
WLP SDSY 4
WY1728 4 
WY1203 5 
WY4766 5 

total vials 98

*edited to add 1203 and 4766


----------



## mofox1

Nice stash! I'm probably sitting on around 30 or so, in about 10 different strains.

An spreadsheet inventory is a good idea - my 'gut feel' for how much/what type grain/hops/yeast I have around has not always proved effective come brew day. :lol:


----------



## kcurnow

Damn that's impressive Yob.
I tried to revive one of my west coast IPAs over the weekend and I got nothing going on. Little yeasties did not survive. I will pull another out and see how it goes.


----------



## Yob

are you thawing quickly? what are your steps? Matt was having trouble too but he was just chucking them into 2lt flasks which is a bit much for the poor devils.

what percentage glycerine?

I drop mine in a glass of tap water and change after 5 minutes and go 500ml > 2000ml/3000ml (depending on the planned brew and the activity)


----------



## Camo6

Ooops, so you're supposed to thaw quickly?
I've got two small starters going atm. My first attempt with frozen vials. I just let the vials thaw in the fridge. The 1217 has started but the 1275 is doing nothing. Might grab another and try again.


----------



## Yob

I was doing the same initially cam, reading suggests that a quick thaw helps avoid shearing of the cell wall during the thaw, anecdotally, I've seen improvement in behaviour since adopting the water thaw approach. Some folks even hold it in their hands till it gets to temp, I reckon that's a bit of overkill though.


----------



## mofox1

Pocket.

Bit cold to start off though.


----------



## kcurnow

Yob said:


> are you thawing quickly? what are your steps? Matt was having trouble too but he was just chucking them into 2lt flasks which is a bit much for the poor devils.
> 
> what percentage glycerine?
> 
> I drop mine in a glass of tap water and change after 5 minutes and go 500ml > 2000ml/3000ml (depending on the planned brew and the activity)


So I chucked the vial in the fridge to defrost overnight then into the 100ml starter then 500ml then 2000ml. My % glycerin is the same as yours as I do a 50/50 water glycerin solution and half fill the 50ml vials pressure cook and cool then top up with the yeast solution. I have then put straight in the freezer. To chill as quickly as possible. Should I be giving the yeast time to absorb the glycerin or is it fine just going straight into the freezer. I will try a fast defrost of another vial this arvo and see if I have any better luck with viability.


----------



## Yob

I go into the fridge for a day or two then freeze, exactly for that reason, right / wrong dunno, just my method


----------



## kcurnow

Well here goes take two. WY1217 in 500ml after the frozen yeast vial was defrosted in a glass of water.


----------



## Camo6

I was in a rush so warmed up a vial of 1469 and tossed it straight in with the 1275.
The 1469 was foaming before it even defrosted! Love that yeast.


----------



## Yob

Brewnut said:


> Well here goes take two. WY1217 in 500ml after the frozen yeast vial was defrosted in a glass of water.
> 
> 
> 
> ImageUploadedByAussie Home Brewer1428295466.560298.jpg


thats quite a vortex mate :beerbang:


----------



## Yob

Transfer station in the fridge as mentioned above


----------



## Camo6

Haha! Transfer Station or as its known in some circles: the Egg Compartment.


----------



## Yob

Only by people without a transfer station...


----------



## Camo6

Bwahahaha!


----------



## mofox1

Oh you(se)!

You crack me up. But leave the eggs outta this - they got enogh problems.


----------



## Yob

A pictures worth a thousand pitches.. or something..


----------



## Alex.Tas

How are you going to cope when you have a bank of each strain? 
Your next "hobby" will have to involve several acres and barley/wheat/rye!


----------



## Yob

Alex.Tas said:


> How are you going to cope when you have a bank of each strain?
> Your next "hobby" will have to involve several acres_* of hops*_!


FTFY 

TBH, I think Ive already peaked with most strains I use for the styles I like.. I need a quantum shift in tastes to further it..

Maybe I'll brew a Lager....


----------



## tateg

So my wife took out 10 vials of yeast last night, while getting something else out. 
Of course she forgot to put the back in the freezer, they been sitting outside all night. 
Now what to do, can I refreeze or is that just a waste of time?


----------



## Yob

That's not great for them but won't kill them outright.

All the same or different?


----------



## tateg

Yob said:


> That's not great for them but won't kill them outright.
> 
> All the same or different?


[SIZE=11pt]Three different strains, mostly house strains. They had only been in the freezer for 3 weeks.[/SIZE]
[SIZE=11pt]So you think it may be worth refreezing them ?[/SIZE]
[SIZE=11pt]i guess i may aswell then test one after a week or so [/SIZE]


----------



## Yob

Personally, I'd chuck them in small starters, just a little jar or something, then Re freeze once they've settled out

one of each that is, that way ypu have a viable fallback if they start to fail


----------



## Danwood

I stepped up, split into 5, and froze a pack of Bry97 (50% viability), just to try my hand.

Using Wolfy's method, and quickly thawed after a short spell in cryo-sleep (2 weeks), it's kicked on like a good 'un in a Citra pale.

Stepping up a whole pack in a 2L starter, then splitting 5 ways is going to result in a decent sample per vial (anyone want to run that through Mr Malty? I had it on android but not on this shitbag Apple phone).

The vial then went through 200ml, 500ml and 2L starters.

Anyway, exciting times indeed. 

Thanks for all the helpful posts.


----------



## menoetes

I'm currently stepping up a Wyeast Kolsch yeast I tried freezing 3 months ago... I say _'tried'_ because it didn't freeze. Maybe my glycerin/water ratios were off but it went into a freezer surrounded by ice but remained in liquid form for the whole time it was in there, has anyone else had an experience like this?

Anyway I through it into a small starter on the stirplate on Wednesday and after 24 hours there was no activity so I lost heart and turned it off. However this morning when I got up, almost 48 hours after pitching it, there's a sign of life in my starter after-all! So I turn the stirplate back on and it forms a krausen almost immediately.

My only concern is the slow start-up and I'm worrying it might be an infection rather than the yeast. Again; has anyone else had an experience like this? I'm just looking for a second opinion before I throw this uncertainty into 21lts of Kolsch wort...


----------



## Mardoo

The not freezing would be high glycerin/water ratio. I often have frozen yeast show little to no movement in my first step with 1.020 wort, which I learned from Yob. The next step everything kicks off as normal. 

I do 3 steps: 150ml at 1.020; 500ml at 1.040; and 2 or 3 litres at 1.040.

Taste the starter wort and that should give you an idea regarding infection. I always taste my starter wort.


----------



## mofox1

Been going yeast freeze crazy over the past couple of weeks.

Just starters to build up stocks of older / limited stock, but I've done:

7 x 15ml vials of WLP023 Burton Ale - 5th gen, same "house" strain that has so far reached 83% AA with the ~1.100 bummock wort.
10 x 15ml vials of MJ Bavarian Wheat - pack was 1 yr past bb - still went nuts. Pack was pitched into a 3L starter, the 10 vials were from 2/3 of the resultant yeast. Other 1/3 built back up to pitch into a wheat.
? x 15ml vials of WLP041 Pacific Ale - last step for this one running now... probably didn't need it. Will easily get 10+ vials with probably enough left over to pitch straight into a PA.

Got stocks of Cal Ale, London Ale & Sth German Lager to build up next. If I'm not sick of it, I've got some older 007, 009 & coopers to revive too.

Safe to say I'm using up a wee bit of DME right now. Fun times.


----------



## lael

ok, so what is the current recommended method for this? 

freezing: 50ml vial - 50% glycerine, 50% decanted yeast slurry (crash cooled, clear wort discarded, slurry packaged). Place in fridge for 2 days before placing in freezer

re-activating: place in room temp glass of water for 5 mins, change water. Sit to bring to room temp OR: hold in hand until hits room temp. Pitch into 200ml @ 1.020, repitch to [email protected] 1.040, repitch to 2L @ 1.040. 

Questions: 
freezing: some mention that 50/50 doesn't freeze and won't show action in the first step. That shouldn't be a problem as long as they are kept at freezing temps right? Might it be better if the cell walls don't freeze solid?
If not - increasing the ratio of yeast slurry to 60/40 or 70/30 should do the job? 

re-activating: Are you guys cold crashing between steps or just adding the 200ml to the 500ml of fresh wort etc?

Did the Melbourne bulk buy of glycerine happen a while back?

I've contacted some suppliers in Sydney and generally they are cosmetics companies. They buy the glycerine in 250Kg packages (that are food grade / organic / BP) and re-pack it, but their facilities/ procedures don't meet/ aren't certified to food grade standards and therefore they can't sell it as such. One supplier I'm thinking of buying from uses a clean room, packs one product at a time, personnell use disposable protective gear (gloves, hair nets etc) use disposable equipment for repacking or wash it between uses with soap and warm water. Thoughts?


----------



## Mardoo

You should absolutely be autoclaving/pressure cooking vials with your glycerine/water yeast medium so if the glycerine is not hermetically packed it shouldn't be such an issue. I haven't had issues yet.


----------



## Yob

so 2 days ago, I took out a vial of 1728 from the deep freezer, this vial has been rock solid for a year or so..

had about 10ml yeast solids in it, I put that into a 500ml 1.020 starter overnight, the next morning, there was visible signs of activity, condensation on the inside of the flask, a lightening of the wort.. so that evening, I made 2 x 2lt starters of 1.040 and split the 500ml between them.. again, overnight they were showing signs of activity, at 18hrs to 24hrs they were starting to foam so Ive turned them off and will let them ferment out before pitching to the Bummock Ale (~1.100 Scotch Ale / Heavy Wee Heavy)

Stoked and pretty impressed at the revival


----------



## Yob

lael said:


> ok, so what is the current recommended method for this?
> 
> freezing: 50ml vial - 50% glycerine, 50% decanted yeast slurry (crash cooled, clear wort discarded, slurry packaged). Place in fridge for 2 days before placing in freezer
> 
> I make up a %50 solution and add my yeast to that so I end up with a ~%25 solution final with solids
> 
> re-activating: place in room temp glass of water for 5 mins, change water. (Yes) Sit to bring to room temp OR: hold in hand until hits room temp. Pitch into 200ml @ 1.020, repitch to [email protected] 1.040, repitch to 2L @ 1.040. (I ALWAYS start with a 500ml @ 1.020)
> 
> Questions:
> freezing: some mention that 50/50 doesn't freeze and won't show action in the first step. That shouldn't be a problem as long as they are kept at freezing temps right? Might it be better if the cell walls don't freeze solid? (anecdotally no, I prefer a less than %50 solution)
> If not - increasing the ratio of yeast slurry to 60/40 or 70/30 should do the job?  (as above, I make up a 50/50 solution of glycerine and sterile water and add that to my vials with 25ml yeast solids/wort)
> 
> re-activating: Are you guys cold crashing between steps or just adding the 200ml to the 500ml of fresh wort etc? (I'll make up the next step to include the volume.. so, if I have a 500ml starter, Ill make up a 200g/1.5lt starter and add the 500ml to make the full 2000ml if that makes sense?)
> 
> Did the Melbourne bulk buy of glycerine happen a while back? (Yes, Mardoo is the man for those details)
> 
> I've contacted some suppliers in Sydney and generally they are cosmetics companies. They buy the glycerine in 250Kg packages (that are food grade / organic / BP) and re-pack it, but their facilities/ procedures don't meet/ aren't certified to food grade standards and therefore they can't sell it as such. One supplier I'm thinking of buying from uses a clean room, packs one product at a time, personnell use disposable protective gear (gloves, hair nets etc) use disposable equipment for repacking or wash it between uses with soap and warm water. Thoughts?  (I do sometimes, if I can be arsed, microwave/autoclave some vials in advance, sometimes my practice is a little less than perfect... Ive not yet sneezed into a vial though)


----------



## lael

How far in advance have you guys pressure cooked the glycerin? Is that - mix water and glycerine and fill vials to half way and then turn the pressure cooker on? For how long? How much water do you put in the bottom of the pressure cooker?


----------



## Mardoo

You can do the glycerine/water vials in advance and keep them in the fridge for a couple weeks. Remember to bring them back to room temp if you're pouring room temp yeast in.


----------



## MartinOC

lael said:


> How far in advance have you guys pressure cooked the glycerin? Is that - mix water and glycerine and fill vials to half way and then turn the pressure cooker on? For how long? How much water do you put in the bottom of the pressure cooker?


Each cooker will be slightly different, so it depends on the model you have. If you can put your vials into a cage or perforated pot to keep them off the bottom, all well & good. Leave the lids on the vials, but loosen them off.

You only need enough water to create the steam under pressure (that's about 120-ish degrees C), so 1cm or so will be heaps. Bring it up to pressure, leave it for 10-15 minutes, then turn the heat off & let it cool down naturally. Overnight is fine.

Once cool, you can open the cooker & screw-down the lids on the vials. They can then be refrigerated or frozen as-is for when you want to transfer & store your yeast (obviously, both the vials & yeast should be at the same temperature when you do this!). Then freeze them.

As an anecdote, I created slants in screw-top McCartney bottles in much this same way & promptly forgot about them for about 10 years :blink:. Not refrigerated the whole time. When I found them again, not a single one was contaminated.


----------



## Bridgey23

Man I wanna freeze yeast and have everything except a pressure cooker. Do I need to pressure cook? Or can I just clean and sanitise?


----------



## Yob

You can use water that's been boiled 3 times over 3 days... Considered sterile at that point..

It's a bit if overkill to be honest, I used just boiled water for rinsing for ages, never noticed any issues, know the risks and be comfortable with the level of risk you choose.

We do tend to err on the side extreme caution most times..


----------



## MartinOC

Bridgey23 said:


> Man I wanna freeze yeast and have everything except a pressure cooker. Do I need to pressure cook? Or can I just clean and sanitise?


Cleaning & sanitising is fine & you'll still get good results. Using a pressure cooker is going that extra step to ensure you get the best results possible every time.

Edit: And it's for yeast-freezing geeks.... h34r:


----------



## Bridgey23

Cheers Yob. I'm new to this. All this! Brewed can goop and sugar about 20 yrs ago. Drinkable but never followed through. Have been inspired lately. Love real beer and jumped straight into all grain and lovin it. Only 5 brews in but remarkably better beer. 
I'm a tight arse like most home brewers and have started splitting liquid yeast starters but see this as an even better solution to yeast costs and of course having particular strains on hand. 
Cheers again Yob you are wealth of information. For us newbies.

Edit: cheers Martin I'm getting geekish. The advice on this forum is fantastic and very much appreciated. I've never found a hobby that has grabbed me like this. Bloody expensive but extremely satisfying.


----------



## Bridgey23

And Yob. If your doing the boil over three days for rinse water method do you wack a lid on it between boils? Put it in the fridge? What's the process?


----------



## mofox1

Bridgey23 said:


> I'm a tight arse like most home brewers...
> 
> ...I've never found a hobby that has grabbed me like this. Bloody expensive but extremely satisfying.


LOL. This.

Pretty much sums up what happens when you slip down that slope..


----------



## Yob

Bridgey23 said:


> And Yob. If your doing the boil over three days for rinse water method do you wack a lid on it between boils? Put it in the fridge? What's the process?


Gladwrap if you can be arsed.. I gave up on it pretty quickly


----------



## Tropico

Would the White Labs vials be suitable for the pressure cooker and freezing?

Edit: Used, empty ones of course.


----------



## sstacey

Do the white labs vials have recycle numbers on them? If it has a 5 in the triangle then it is polypropylene and resistant to pressure cooking. However, sometimes tube lids are made from different plastics to the main body, so you need to make sure the lid is also heat resistant. Please let me know as I also keep yeast and if the white labs vials can be used for slants it would be ideal (so far I have only used wyeast).


----------



## coopsomulous

The white lab vials are unblown 1.25L PET bottles. I wouldn't use them for slants as they would soften quickly at high temps.

I have seen the new White Labs packaging for WLP001 at a local HBS recently, so I would assume that the time for the PET tubes are nearly over.


----------



## Yob

You can use them, they soften but harden up again, the biggest pain in the Arse is they don't stand up though so are awkward in the freezer.


----------



## Tropico

No PET number on the vial and the PET bottles have "1" in the triangle with "PETE" underneath.

I'll try boiling one first, to see how soft it gets.


----------



## Whiteferret

Yob said:


> so 2 days ago, I took out a vial of 1728 from the deep freezer, this vial has been rock solid for a year or so..
> 
> had about 10ml yeast solids in it, I put that into a 500ml 1.020 starter overnight, the next morning, there was visible signs of activity, condensation on the inside of the flask, a lightening of the wort.. so that evening, I made 2 x 2lt starters of 1.040 and split the 500ml between them.. again, overnight they were showing signs of activity, at 18hrs to 24hrs they were starting to foam so Ive turned them off and will let them ferment out before pitching to the Bummock Ale (~1.100 Scotch Ale / Heavy Wee Heavy)
> 
> Stoked and pretty impressed at the revival
> 
> 1728.JPG


It wouldn't happen to be the one I left for you when you hosted the swap would it?
If so good to know about the ones I've got still.


----------



## Alex.Tas

I use bottled water for top up water rather than cooled boiled water. It should be pretty sterile shouldn't it?


----------



## Yob

whiteferret said:


> It wouldn't happen to be the one I left for you when you hosted the swap would it?
> If so good to know about the ones I've got still.


God only knows mate. Fired like a demon though, letting them ferment out now.... 

Win


----------



## Whiteferret

Was it a different vial with a white top?


----------



## Yob

Na mate, blue lid


----------



## takai

Ive been considering using the baby stuff steriliser that we have for sterilising. Its a steam based unit, over 15mins or so. Anyone use something like that?


----------



## lael

Anyone considered making a gel like freezing container?

http://www.spine-health.com/blog/how-make-your-own-gel-ice-pack-or-moist-heat-pack


----------



## Bridgey23

"Ive been considering using the baby stuff steriliser that we have for sterilising. Its a steam based unit, over 15mins or so. Anyone use something like that?"
Great idea sounds like a winner unless someone can come up with a reason not to!
Something like this?
http://www.ebay.com.au/ulk/itm/191698474314

Edit: couldn't work out how to copy takai's post like everyone else seems to manage when replying.


----------



## Bridgey23

Lol think I worked it out. Quote button yeah?


----------



## Tropico

Well I gave the old White Labs vial a boil for 5 minutes or so. Definitely became soft, around the screw top more so than the main body.

Had a bit of a search too, apparently PETE starts to soften at ~80C and becomes softer as temperature increases until it becomes liquid ~240C.

The black screw-top faired much better than the clear vial which became slightly opaque, either from the chlorine or the label adhesive or both.

Maybe leaving the black screw-top loosely on in a pressure cooker might help retain the shape.

Found a pressure cooker that is also a slow cooker and also does yogurt. SWMBO would love one of them, I might buy her one.

EDIT: As a special present for her to show my appreciation.


----------



## MartinOC

Not gonna do the multi-quote thing here..

Second-hand pressure cookers can be had for nix (check your grandmother's stash of stuff she's collected over a lifetime). Maybe all that's needed is a new sealing ring. I picked one up from a local "pressure-cooker repair" place for $10. 'Been using it for years.

To get proper sterilisation, you need to get the contents to ~ 120C for 10-15 minutes. Job done.

I've been eyeing-off one of these babies for awhile, but will wait until I can justify the cost to SWMBO: http://www.ebay.com.au/itm/Presto-21-litre-Pressure-Canner-Cooker-6-Ball-Mason-Quart-Preserving-Jars-/151710568294?hash=item2352a78f66


----------



## GalBrew

Tropico said:


> Would the White Labs vials be suitable for the pressure cooker and freezing?
> 
> Edit: Used, empty ones of course.


Nope they are PET, not polypropylene.


----------



## Tropico

MartinOC said:


> Not gonna do the multi-quote thing here..
> 
> Second-hand pressure cookers can be had for nix (check your grandmother's stash of stuff she's collected over a lifetime). Maybe all that's needed is a new sealing ring. I picked one up from a local "pressure-cooker repair" place for $10. 'Been using it for years.
> 
> To get proper sterilisation, you need to get the contents to ~ 120C for 10-15 minutes. Job done.
> 
> I've been eyeing-off one of these babies for awhile, but will wait until I can justify the cost to SWMBO: http://www.ebay.com.au/itm/Presto-21-litre-Pressure-Canner-Cooker-6-Ball-Mason-Quart-Preserving-Jars-/151710568294?hash=item2352a78f66


Stuff the cost, SWMBO is worth it


----------



## Budron

Hey Takai, 

I use this for my yeast harvesting and haven't had any problems so far. Go for it!


----------



## Tex N Oz

I've not read this whole thread but was wondering if anyone has mentioned that growing your cryo-culture in a very nutrient poor medium will greatly increase viability.
the cell walls of well fed yeast have a high tendency to rupture leaving only those colonies that are less than optimum because of pre-existing nutrient uptake issues.


Edit: I should elaborate considerably more. The yeast needs to be cultured @25°C for 72h in YM media.
Grown in 10ml vials.

The YM media recipe is such:

3g yeast extract
3g malt extract
5g peptone
10g glucose
deionised water to make to 1 litre

The cryoprotectant solution should be 10%v/v glycerol mixed equally with the innoculum


----------



## Yob

No..

Cmon Michael Burton aka digital stirplates with the temp controlled unit though...


----------



## lael

Tex N Oz said:


> I've not read this whole thread but was wondering if anyone has mentioned that growing your cryo-culture in a very nutrient poor medium will greatly increase viability.
> the cell walls of well fed yeast have a high tendency to rupture leaving only those colonies that are less than optimum because of pre-existing nutrient uptake issues.
> 
> 
> Edit: I should elaborate considerably more. The yeast needs to be cultured @25°C for 72h in YM media.
> Grown in 10ml vials.
> 
> The YM media recipe is such:
> 
> 3g yeast extract
> 3g malt extract
> 5g peptone
> 10g glucose
> deionised water to make to 1 litre
> 
> The cryoprotectant solution should be 10%v/v glycerol mixed equally with the innoculum



What is YM media?


----------



## Mardoo

And can you provide some links so we can learn more about this?


----------



## Tex N Oz

lael said:


> What is YM media?


YM media is a product produced by Beckton Dickinson. It's like calling facial tissues a Kleenex of sorts. 



Mardoo said:


> And can you provide some links so we can learn more about this?


Most of it's from papers I've collected over the years. I used to be hardcore into lyophilization.
I could try to scan in some shit.

I've been toying with this for years.


----------



## Tex N Oz

Bad news for everyone though.. without proper lyophilization, none of these innoculums will be viable past 2 years with the temperatures that you're using.
You don't even start to get into cryo-preserving until -80°C.
You might get some petite colonies on your strikes but you don't want those. They're the fucked ones that are just barely hanging on and very hard to start. You'll also notice a HUGE change in profiles as mitochodrial damage is usually the cause.


----------



## Tropico

No shit


----------



## Yob

so not really useful at all.. I keep mine at a pretty steady -20'c and am happy with the results thus far without it being overly complicated...

one of the things that got me into this side of the yeast ranching game over many other methods.

KISS


----------



## Bridgey23

Looks like your ganna have to look into liquid nitrogen storage now Yob. Does it ever end?[emoji6]
Keep us informed if you do!


----------



## Bridgey23

Ha ha how far do you wanna take it?
http://www.boconlineshop.com/shop/en/au-boc-industrial-store/liquid-nitrogen-storage/mve-lab-20-liquid-nitrogen-dewar-mve13492631-p


----------



## Tex N Oz

Yob said:


> so not really useful at all.. I keep mine at a pretty steady -20'c and am happy with the results thus far without it being overly complicated...
> 
> one of the things that got me into this side of the yeast ranching game over many other methods.
> 
> KISS


It's important to understand that raising yeast from a cryo or lypho state isn't as simple as pitching into some wort and off it goes. You must ALWAYS strike your sample first, then grown it on a petri dish and then start a 10ml starter then up from there. There's no easy way to start from a dehydrated sample. Even those samples that are frozen in a freezer are considered dehydrated.
The yeast have many many colonies and you want to start only the healthiest of these and not a mix of these.
If your goal is to bank yeast that will keep for years and years and years at 1°C, then you must involve lyophilisation or true cryo-freezing (liquid nitrogen @ - 160°C max) and then understand that it will take you upwards of 2 weeks to have a viable starter.
I truly hate to come across as a "know it all" and sometimes I come across that way but I have extensively studied this and only sharing my experiences and learning.
There's a reason you don't buy frozen yeast in the freezer section of your LHBS.


----------



## Yob

Practical experience suggests different. 

Results are speaking for themselves. Are there damaged colonies? Probably, does it make a noticeable difference? No.

DIGAF? No, the system I utilise works well enough and I'm happy with it and that's the important thing here.. It's a simple, effective, home brew solution to a yeast bank that works. 

Perhaps you should start another thread on that method so as not to De rail this one.


----------



## Tex N Oz

Yob said:


> Practical experience suggests different.
> 
> Results are speaking for themselves. Are there damaged colonies? Probably, does it make a noticeable difference? No.
> 
> DIGAF? No, the system I utilise works well enough and I'm happy with it and that's the important thing here.. It's a simple, effective, home brew solution to a yeast bank that works.
> 
> Perhaps you should start another thread on that method so as not to De rail this one.


That's fair enough. How long are you able to keep viable samples?


----------



## Yob

Well over 18 months currently... And counting


----------



## Tex N Oz

Yob said:


> Well over 18 months currently... And counting


Are you finding that it works better for some strains than others?


----------



## Yob

I'm finding the glycerine content is more critical, I went from %15 to %50 but have found the best results at about %25-30


----------



## Tex N Oz

That's interesting. All of my experience has been with cryo freezing @ 5% total solution.
I might give the higher concentration and higher temperatures a try. There are a lot of times it would be handy to have a larger sample ready to go.
Have you given any of the lager yeasts a go?


----------



## Yob

I've got some frozen but not Re cultured, I tend not to brew a lot of lagers. I'll pull one out this week just for shits and giggles


----------



## Tropico

According to Chirs White and Jamil Zainasheff, it is possible to have a reliable shelf life up to 2 years and maximum shelf life longer than 5 years when stored between -2 and -19C. At -80C , they quote both as "indefinite".


----------



## menoetes

Just a quick follow up from my post about a week back; the Wyeast Kolsch slurry/glycerine solution that didn't actually freeze in the freezer did kick off in the end (also I'm quite sure it's not an infection like I first feared) and I stepped in up from 100ml to 400ml then 1lt then 2.4lts - nice and easy.

It's found it's way into my Kolsch wort last night and I have high hoped for it. So for future reference - even if your yeast slurry doesn't freezer, hold onto it - it might still make a go of it if treated gently in restarting.


----------



## barneey

Following Yob's method, Green Belt yeast frozen Dec 14, resurrected October 15 so I would call that a success. 

Cheers


----------



## Yob

My 1728 that's been frozen well over a year, probably 18 months has taken my scotch ale from ~1.100 to 1.020 in just on 5 days.

I'm gunna take that as a viability and vitality win also.


----------



## idzy

Yob said:


> My 1728 that's been frozen well over a year, probably 18 months has taken my scotch ale from ~1.100 to 1.020 in just on 5 days.
> 
> I'm gunna take that as a viability and vitality win also.


Like a cut cat.

Just put down 4 vials of Nottingham with 35% glycerine.


----------



## michaeld16

What are poeple thoughts on freezing a pack of roeselare blend? Has anyone tried it, would the bugs survive? I may just have to give it a go and see


----------



## Danwood

I don't think it'd be any different. You're still adding the same glycerine buffer to stop the same (same basic structure, anyway) cell walls from being damaged in the freezing process. 
That's my understanding of it.
I'm stepping up and splitting a Wyeast 3031 for freezing (saison/brett) currently and predict the same excellent results as I've had with my regular yeasts.


----------



## michaeld16

Sorry if this is covered in the thread already, but do ya need to decant off the glycerine solution once defrosted and then put it in a starter might be a dumb question but i only thought of it after pitching the whole vial in a 200ml starter last night not seeing any action yet, this is my first attempt at reviving my frozen stock.


----------



## Mardoo

No, but you can. First step often shows precious little action in my experience. Next ones are as normal.


----------



## michaeld16

Awesome thats good news, yeah i wasnt expecting much action on the first step will step up to a 500ml tomorrow night


----------



## Yob

Go to 2000 if you have it, x10 step is good, I'll often go into 500 ml off the bat and then 2000ml


----------



## MartinOC

Yob said:


> Go to 2000 if you have it, x10 step is good, I'll often go into 500 ml off the bat and then 2000ml


Erm...unless I'm missing something that's a x4 step (in volume).

Have you been sniftering the Bummock a bit early there, Yob?


----------



## Yob

im special


----------



## Yob

MartinOC said:


> Erm...unless I'm missing something that's a x4 step (in volume).
> 
> Have you been sniftering the Bummock a bit early there, Yob?



just for clarity, Michael is proposing he pitch his 200ml starter into a 500ml starter, Im suggesting he go to a 2000ml starter..

What I generally do is a different case again...


----------



## michaeld16

All clear here will get a 2l starter going tomorrow after work


----------



## michaeld16

So the next morning after pitching the 200ml into 2l of 1040 wort it was going off smells like success. Big thanks to the contibuters of this thread this means liquid yeast whenever i need it.


----------



## tateg

Quick question, how / what is the best way to store the tube in a frost free freezer ?


----------



## Yob

Packed with salt ice in an insulated container of some sort just to minimise temp swings


----------



## tateg

thanks yob
do u mean make the salt ice ? i was thinking maybe a small foam esky but haven't been able to source one


----------



## Yob

Yep, salt ice will be colder, I initially used a cluplock box and gaffe taped Styrofoam to it... Worked but I ended up dedicating space in the deep freezer... Better


----------



## mofox1

Yeh... I use a 3 or 4 ish litre capacity styrofoam container, which I believe just came from Coles or Woolies. Enough for ~100 vials (mixed 50ml & 15ml).

Basically looks like a tiny nude esky. :lol:


----------



## VP Brewing

This thread has inspired me to make these up at work today. I'm picking up 3 Wyeast smack packs on Monday so will split them up. 
Living 2hrs north of Melb is a pain in the arse when it comes to getting yeast sent up in good condition, especially in summer. 
They are 28ml containers with 6ml of water and 6ml of glycerine. Will put about 12ml of yeast in each one so hopefully get about 8 per pack. 
Autoclaved at 121°C for 25min.


----------



## VP Brewing




----------



## kcurnow

VP Brewing said:


> ImageUploadedByAussie Home Brewer1446185281.580852.jpg


now thats a steriliser!!


----------



## MartinOC

VP, that is a thing of beauty & you're definitely well on the track for success.


----------



## Yob

a peak inside the freezer reveals...





addiction


----------



## tateg

Yob said:


> Yep, salt ice will be colder, I initially used a cluplock box and gaffe taped Styrofoam to it... Worked but I ended up dedicating space in the deep freezer... Better


Just how much salt in the water are we talking ? Simple as just crush and place abound the vials?


----------



## VP Brewing

Yob said:


> a peak inside the freezer reveals...
> 
> 
> 
> 
> 
> Vials.JPG
> 
> addiction



So many choices!


----------



## Yob

tateg said:


> Just how much salt in the water are we talking ? Simple as just crush and place abound the vials?


Make a bunch of ice cubes


----------



## MartinOC

I'll see your selection & raise you just a couple of the bags I just pulled from my freezer:




All opened once only to decant excess liquid & add glycerine.

All packaged within BB-date. Free.


----------



## Mardoo

Careful folks, low-swingin' dick around here


----------



## danestead

Have any of you guys checked viability of your frozen yeast with a microscope? I just had a vial of mine checked and it revealed 99.9% dead and munted yeast. Very few live cells.

Wyeast 1217 West Coast IPA
25% glycerine, 25% water, 50% wyeast poured straight from a 2 week old packet
Stored at -20 degrees
It was in solid form however I think it had liquidised at some stage because layers are now visible
1 year in the freezer


----------



## MartinOC

High-swinging if you don't mind.......


----------



## mofox1

danestead said:


> Have any of you guys checked viability of your frozen yeast with a microscope? I just had a vial of mine checked and it revealed 99.9% dead and munted yeast. Very few live cells.
> 
> Wyeast 1217 West Coast IPA
> 25% glycerine, 25% water, 50% wyeast poured straight from a 2 week old packet
> Stored at -20 degrees
> It was in solid form however I think it had liquidised at some stage because layers are now visible
> 1 year in the freezer


Great feedback Dano... did you have any samples with a higher glc % that you could get checked?

I usually go around 30 - 35% glc, samples are usually only-just-not-quite-frozen at these levels... the whole idea about adding glycerine is to be able to reduce the temp (and therefore the attrition rate with higher temps) while not subjecting your yeast buddies to cell wall destroying water crystallization death (there is a word for when then cell wall is ruptured and the contents are spilled out, but after eleventy or something beers I can't remember it).

I haven't got it right with all of them, probably about half still look "frozen" which is probably why that first starter step is so laggy and uneventful... would be handy to see if there is an increase in viability (and observations on cell health / mutations) with higher glc rates.


----------



## danestead

mofox1 said:


> Great feedback Dano... did you have any samples with a higher glc % that you could get checked?
> 
> I usually go around 30 - 35% glc, samples are usually only-just-not-quite-frozen at these levels... the whole idea about adding glycerine is to be able to reduce the temp (and therefore the attrition rate with higher temps) while not subjecting your yeast buddies to cell wall destroying water crystallization death (there is a word for when then cell wall is ruptured and the contents are spilled out, but after eleventy or something beers I can't remember it).
> 
> I haven't got it right with all of them, probably about half still look "frozen" which is probably why that first starter step is so laggy and uneventful... would be handy to see if there is an increase in viability (and observations on cell health / mutations) with higher glc rates.


Unfortunately no I havent got any with any different glycerin contents. Ive only done the 1 batch.

My freezer used to be set on -15 deg but the vials were liquid at that setting so I upped it to -20 to freeze them solid with the sole purpose of stopping the settling out and layering of yeast. Why? I dont know. Just because. Maybe liquid form would of been better at -15 however in this thread I think people have been successful with both liquid and solid versions.

Id really like to hear some more scientific evidence of viabilities rather than yes it fired up quick etc etc.


----------



## Yob

What's the process you use to freeze? Do you let them sit in the fridge for a few days prior to the deep freeze?

My ratio's are roughly the same and there's no way I get that much death, the starters fire up too fast.


----------



## Midnight Brew

danestead said:


> Have any of you guys checked viability of your frozen yeast with a microscope? I just had a vial of mine checked and it revealed 99.9% dead and munted yeast. Very few live cells.
> 
> Wyeast 1217 West Coast IPA
> 25% glycerine, 25% water, 50% wyeast poured straight from a 2 week old packet
> Stored at -20 degrees
> It was in solid form however I think it had liquidised at some stage because layers are now visible
> 1 year in the freezer


How did you freeze your yeast? Did you add straight to the freezer? Did you bring them down to below zero then freeze?

I ask this because a fellow brewer, Grainer, pointed me in the right direction to freeze as quickly as possible. Ice bath of salt water and throw the vials in works for me.

First batch I froze I slow froze them and killed them all. I tried Grainer's ice bath suggestion and it worked and those yeast are still viable 18+ months later. (I use a 15% Glycerine solution)


----------



## danestead

Midnight Brew said:


> How did you freeze your yeast? Did you add straight to the freezer? Did you bring them down to below zero then freeze?
> 
> I ask this because a fellow brewer, Grainer, pointed me in the right direction to freeze as quickly as possible. Ice bath of salt water and throw the vials in works for me.
> 
> First batch I froze I slow froze them and killed them all. I tried Grainer's ice bath suggestion and it worked and those yeast are still viable 18+ months later. (I use a 15% Glycerine solution)


I can't actually remember 100% how I froze them however I do remember reading that freezing them as quickly as possible is beneficial. I would take a guess and say I did an ice bath and then put them in the freezer. I also remember setting my freezer to its max of -30 for the first day, then let it go back to its normal setting of -20.

Having said that, I think I've read that guys on here like Yob fridge them for a day then freeze them.

I guess the main point of my post regarding the 99.9% fatality of my frozen yeast is to find out who has actually taken a more scientific approach like looking at the yeast under a microscope to see what the state of them are. Considering the fatality rate my batch has had, I don't think that it satisfies me enough anymore to assume other peoples yeast have great viability and health because they have said their starters are firing up well and attenuating as per normal. I want to see the results of their freezing attempts under a microscope before I go ahead and freeze more yeast.


----------



## Yob

Bring round the microscope. I'm interested now myself


----------



## kcurnow

here is my microscope, complete with 5mp lens camera. I have the haemocytometer just need to get J-count (automated cell counting software) loaded on a computer and then get some time to fire up one of my frozen batches and do some viability tests. As well as move to QLD. Sooooo it might be a while.


----------



## Yob

So can I bring a vial on Tuesday?


----------



## Mardoo

Selling the mic???


----------



## kcurnow

Mardoo said:


> Selling the mic???


 nope!!


----------



## kcurnow

Yob said:


> So can I bring a vial on Tuesday?


hmmm I don't think I'm that organised to do all the dilution, staining and counting just yet.


----------



## Yob

Mkay


----------



## idzy

Pulled some WY3787 out from February 2014 (~19 months), seems to be doing well. From memory was a 35% glycerine, so put glycerine upto 17ml mark and then filled to top of 50ml with decanted starter.

This is the first one I have started, as I have been freezing, but not needing to re-use the same stains. I was a bit worried because it had a settled solid section on the vial, but it wasn't frozen and a few shakes and the settled section mixed with the rest. Put it into 400ml and then didn't decant and poured onto 800ml of fresh starter wort, due to the yeast type.


----------



## Yob

idzy said:


> Pulled some WY3787 out from February 2014 (~19 months), seems to be doing well. From memory was a 35% glycerine, so put glycerine upto 17ml mark and then filled to top of 50ml with decanted starter.
> 
> This is the first one I have started, as I have been freezing, but not needing to re-use the same stains. I was a bit worried because it had a settled solid section on the vial, but it wasn't frozen and a few shakes and the settled section mixed with the rest. Put it into 400ml and then didn't decant and poured onto 800ml of fresh starter wort, due to the yeast type.
> 
> 
> 
> 20151104_180002a.jpg


What time period Idzy?


----------



## idzy

Yob said:


> What time period Idzy?


First step was 36, current step has been 24.


----------



## michaeld16

How are folks here calculating pitching rates? Say if im using the brewers friend calculator what do you put in the number of liquid yeast packs/vials field? If I split the pack into 4 vials should I just put .4 and maybe back the manufac date back a bit. What works for others here.


----------



## VP Brewing

Just pitched some wyeast 1275 into a brew this arvo. Has only been frozen for a couple of weeks but the smack pack was almost 4 months old when I split and froze it. 
It was 6ml glycerin, 6ml water and ~12ml yeast into a 28ml container. 
Thawed and stepped up into 200ml @ 1.030 then 1.2L @ 1.040. 
Worked first time no dramas thanks to this thread! 
Cheers!


----------



## Yob

michaeld16 said:


> How are folks here calculating pitching rates? Say if im using the brewers friend calculator what do you put in the number of liquid yeast packs/vials field? If I split the pack into 4 vials should I just put .4 and maybe back the manufac date back a bit. What works for others here.


Not a direct answer... Sort of.. But doing several starter steps will get you back to the 100 billion or any other number you need


----------



## Yob

Yob said:


> I made a little foam shielded container today and put my first ones in.. had a look before and they look to be sitting at about -10'c


1.095 >>>>>>> 1.018 @ day day 6 from pitching.... 2 year old yeast...

just sayin.. :beerbang:


----------



## jibba02

Probably all the bacteria in the water eating all the sugars


----------



## Yob

Sure tastes ok... Probably shouldn't have committed the IIIPA to it though... Will just have to drink it quickly...


----------



## mofox1

Yob said:


> Sure tastes ok... Probably shouldn't have committed the IIIPA to it though... Will just have to drink it quickly...


Aaaand the problem being?


----------



## Mattrox

Yob said:


> Sure tastes ok... Probably shouldn't have committed the IIIPA to it though... Will just have to drink it quickly...


pfft... It will be fine. RDWAHAHB


I have been following along this thread for a very long time now. It has convinced me that I MUST freeze yeast. For very minimal outlay and really not much extra work, you get the convenience of having a ready supply of different yeasts on hand ready to whip up into a starter. 

I am going to try with some Coopers yeast. I have made up a starter, but my only glassware is a 1L Moccona jar. I made up 1L of DME wort and pitched the slurry from 5 Pale Ales into it. It got to wpok straight away.

I put the starter in the fridge to chill after 4 days and poured off the spent wort. Because I only have a 1 L jar I made up 1 more litre of DME starter and poured it over the yeast.

Now this is going to have fermented out before I am really ready to brew so I had planned to chill and pour off the spent wort and store for a couple of days. I was going to rouse the yeast in the little bit of liquid left draw off 20ml for freezing and pitch the rest.

Does this sound like something that would work?


----------



## Camo6

Mattrox said:


> pfft... It will be fine. RDWAHAHB
> 
> 
> I have been following along this thread for a very long time now. It has convinced me that I MUST freeze yeast. For very minimal outlay and really not much extra work, you get the convenience of having a ready supply of different yeasts on hand ready to whip up into a starter.
> 
> I am going to try with some Coopers yeast. I have made up a starter, but my only glassware is a 1L Moccona jar. I made up 1L of DME wort and pitched the slurry from 5 Pale Ales into it. It got to wpok straight away.
> 
> I put the starter in the fridge to chill after 4 days and poured off the spent wort. Because I only have a 1 L jar I made up 1 more litre of DME starter and poured it over the yeast.
> 
> Now this is going to have fermented out before I am really ready to brew so I had planned to chill and pour off the spent wort and store for a couple of days. I was going to rouse the yeast in the little bit of liquid left draw off 20ml for freezing and pitch the rest.
> 
> Does this sound like something that would work?


Would it work? Yes but with far more risk to preserving a decent sample. Freezing yeast is cheap and easy but it doesn't hurt to be prepared. If you want cheap starter jars then V8 juice bottles work really well.


----------



## Mattrox

Camo6 said:


> Would it work? Yes but with far more risk to preserving a decent sample. Freezing yeast is cheap and easy but it doesn't hurt to be prepared. If you want cheap starter jars then V8 juice bottles work really well.


What aspect would you improve?

The stepping up........

Or should I take out the sample to freeze in 3 days as soon as it is ready?


----------



## MartinOC

Camo6 said:


> If you want cheap starter jars then V8 juice bottles work really well.


If you want even cheaper, then generic Coles/Safeway passata bottles are the go. They're also great for rinsing-down a yeast cake & storing for future use, as they're tall & thin & you can actually see the stratification in the trub/yeast as it settles.


----------



## Midnight Brew

Greenbelt III lives on!


----------



## Camo6

Mattrox said:


> What aspect would you improve?
> 
> The stepping up........
> 
> Or should I take out the sample to freeze in 3 days as soon as it is ready?


Was just saying that if you go to the trouble of freezing yeast you want to be decently equipped to handle the yeast cleanly. Stepping up a starter from a commercial beer can be a bit risky even before you try freezing it. Not saying it can't be done easily but you might want more than a 1l Moccona jar! I like splitting fresh yeast for the freezer but have occasionally frozen second or third generations (Yob's Greenbelt for example is still in the bottom of the chesty).
Your stepping up method, while not exactly calculated, will grow yeast and is very similar to the approach I was taking before my hiatus from brewing. I'll try and be more clinical when I recommence but it's all so easy to wing it although it doesn't always produce the cleanest ferment. 



MartinOC said:


> If you want even cheaper, then generic Coles/Safeway passata bottles are the go. They're also great for rinsing-down a yeast cake & storing for future use, as they're tall & thin & you can actually see the stratification in the trub/yeast as it settles.


Definitely! I probably should have referenced these as I have more of them than the V8 jars. I mainly use the standard jam jar size as these fit in my pressure cooker but the passata jars are great for separating and I even use them as blow off bottles for the brew buckets when I'm not expecting a krausen explosion.
Will be digging out some frozen samples this weekend for the first brew in almost a year. Will probably bash out some basic smash type beers to use up all the old grain and hops I have and get my bearings. Pretty sure most of my frozen hops are Hop Dealz from as far back as 2012!


----------



## MastersBrewery

Good to hear ya gettin back in the saddle Camo!


----------



## Mattrox

Camo6 said:


> Was just saying that if you go to the trouble of freezing yeast you want to be decently equipped to handle the yeast cleanly. Stepping up a starter from a commercial beer can be a bit risky even before you try freezing it. Not saying it can't be done easily but you might want more than a 1l Moccona jar! I like splitting fresh yeast for the freezer but have occasionally frozen second or third generations (Yob's Greenbelt for example is still in the bottom of the chesty).
> Your stepping up method, while not exactly calculated, will grow yeast and is very similar to the approach I was taking before my hiatus from brewing. I'll try and be more clinical when I recommence but it's all so easy to wing it although it doesn't always produce the cleanest ferment.


Coopers' instructions say use 6 tablespoons of sugar with 600mL of water for yeast slurry from 4 bottles then pitch at active ferment. Now, assuming is dangerous, but I do assume that this is for most of their Kit and Kilo type recipes which typically end up around 1.045 or so. 

So without molar calculations, I used 100g of malt for 1L of water for 5 bottles worth of yeast slurry. But the yeast is going to be pitched on a higher OG wort than 1.045 so being limited in volume, I used the same 100g of malt in 1L of water to "step-up". The big moccona jars hold that easily. And the DIY stir plate worked well and today I had a jar looking like Murray River water. I'll try to calculate the amount of yeast from the "allow the yeast to settle, then pour off the spent wort until equal amounts of yeast and wort remain. Re-suspend the yeast and measure (I'll Calculate) the volume and use the 1B cells per ml" method. 

I also read that 100B cells pitched into a 1L starter on a stir plate will yield 246B cells. So that gives me a ball park to work from. 

We'll see how the yeast works on the wort. And I'll find out a little about freezing yeast too.


----------



## Yob

Coopers give shit advice about yeast


----------



## Yob

Mattrox said:


> Coopers' instructions say use 6 tablespoons of sugar with 600mL of water for yeast slurry from 4 bottles then pitch at active ferment. Now, assuming is dangerous, but I do assume that this is for most of their Kit and Kilo type recipes which typically end up around 1.045 or so.
> 
> So without molar calculations, I used 100g of malt for 1L of water for 5 bottles worth of yeast slurry. But the yeast is going to be pitched on a higher OG wort than 1.045 so being limited in volume, I used the same 100g of malt in 1L of water to "step-up". The big moccona jars hold that easily. And the DIY stir plate worked well and today I had a jar looking like Murray River water. I'll try to calculate the amount of yeast from the "allow the yeast to settle, then pour off the spent wort until equal amounts of yeast and wort remain. Re-suspend the yeast and measure (I'll Calculate) the volume and use the 1B cells per ml" method.
> 
> I also read that 100B cells pitched into a 1L starter on a stir plate will yield 246B cells. So that gives me a ball park to work from.
> 
> We'll see how the yeast works on the wort. And I'll find out a little about freezing yeast too.


pitching such a small amount of yeast into a large starter isnt good for them

pitching stressed, old yeast onto above 1.020 wort as a first step isnt good for them

pitching dregs onto sugar isnt good for them

**** coopers.


----------



## MartinOC

Mattrox said:


> Now, assuming is dangerous
> Assuming IS dangerous
> 
> So without molar calculations,
> Uh-oh! We're getting into "Seriously Geeky" here......
> 
> I'll try to calculate the amount of yeast from the:
> 
> "allow the yeast to settle, then pour off the spent wort until equal amounts of yeast and wort remain. Re-suspend the yeast and measure (I'll Calculate) the volume and use the 1B cells per ml" method.
> 
> I also read that 100B cells pitched into a 1L starter on a stir plate will yield 246B cells. So that gives me a ball park to work from.
> 
> Yeah, a good ballpark. Up or down a little won't hurt.
> 
> We'll see how the yeast works on the wort. And I'll find out a little about freezing yeast too.
> 
> Good idea


Mate, I absolutely admire your quest for perfection in your beers & I share that quest with you......

Sometimes the best thing is just to experiment.......there's the fun..... 

JUST MAKE BEER!!!

Edit: Yob also hit it earlier.


----------



## Yob

^^^ have fun, experiment, just make beer ^^^

well said...

may I add... have LOTS of freezer space.. and not that shitty frost free frezer space either... good old fashioned deep freeze


----------



## Mattrox

I'm using this as a dry run, so my intention was to freeze the yeast and leave it for a month or 2...... which will likely turn into 4 or 6, and see if I can make a starter that ferments and isn't smelly.

By that time I may have procured a conical flask or two.

Next time I'll use a sixer of Pale Ale on less wort at 1.020. If I treat the yeast more tenderly, they might just sleep for me and wake up rearing to go.


----------



## Yob

when you bank your yeast, you want them in as healthy a position as you can get them, its a pretty hostile environment and we arent doing them any favors doing it to them.

a small starter for your dregs, ~100ml max of 1.020 and another of 1.040 at 500ml to get good growth and vitality restoration is essential.

Think of them like bears, they need to be well fed before hibernating.

BY all means.. experiment.. (ed It's rad fun) its what got me into this too and now I have about 150 vials frozen off :lol:


----------



## Mattrox

Looks like 186B roughly. It didn't build up as much as I thought it might. 

Learning curve.


----------



## Yob

So my 2 year old greenbelt on the IIIPA 

1.095 fast forward 6 days.... 1.012

Almost %12 so approaching threshold I'd say.. I'd better get some dry hops in there


----------



## Midnight Brew

Beast!


----------



## CmdrRyekr

What's the go with freezing yeast blends ladies? Have a 3031PC Saison/Brett blend smackpack here I'm planning on growing up and brewing with; but as far as freezing slants goes I guess without a) knowing the ratios of the original blend and b ) without plating it out and splitting the strains, it's probably going to be a bit of a shitshow.

Thoughts?


----------



## jibba02

Don't freeze slants


----------



## Yob

In principle, I don't see why it wouldn't freeze the same as any other yeast with the process outlined in this topic. I certainly wouldn't be freezing slants though. Plating it out kinda defeats the purpose of it being such a simple process, just mix the vial well, split, add glycerine and freeze is what I'd do.

You have a deep freezer? If not there are other things you need to do.


----------



## CmdrRyekr

Yob said:


> In principle, I don't see why it wouldn't freeze the same as any other yeast with the process outlined in this topic. I certainly wouldn't be freezing slants though.


I said slant, but not really what I meant. Vials then.



Yob said:


> Plating it out kinda defeats the purpose of it being such a simple process, just mix the vial well, split, add glycerine and freeze is what I'd do.


I'm wondering if the strains need to be split, cultured, then frozen and re-mixed in the correct ratios to maintain the original blend - hence my other comments.

Sure I can see why you could just freeze a sample as it came out of the pack, but that doesn't account for different growth rates between sacc and brett as in this example.



Yob said:


> You have a deep freezer? If not there are other things you need to do.


No. Isn't the whole premise here that it can be done in a regular freezer we all have at home? Put the vials in a foam box with ice packs to buffer the freeze/thaw cycle from the frost-free bullshit, meaning we don't need a deep freezer.


----------



## Yob

Im lucky in that I have a 1000lt deep freezer, but yes, youll need to insulate, and while that is an acceptable option..if I had a frost free here's what I would do.

take a suitable sized container and fill with salt/water get as many tubes as you can and fill with water and place in the salt water container and freeze, when you need to freeze off a vial, slip one of the water ones out and replace with your vial of yeast.


----------



## mofox1

I use one of these. $9 at aussie disposals.

Dare say you could make your own with a bit of spare styro and gaffa tape that would work just as well.


----------



## Yob

mofox1 said:


> I use one of these. $9 at aussie disposals.
> 
> Dare say you could make your own with a bit of spare styro and gaffa tape that would work just as well.


Im suggesting that if that were filled with a salt water frozen solution it would be less prone to any temperature fluctuation from a that cycle, whereas, if the vials are in there on their own, its the surface of the vials that act as the coolant.. I know which I'd rather the warmer air be attracted to first


----------



## Yob

so Ive decided to tackle storage, my current method is a bit random and it can be hard to find a vial of something in the 55lt container.




there is a fair bit of cutting involved but its going to be ******* stirling..

15 x 11 and I can fit at least 2 layers in the tub which lives in the bottom of my 1100lt deep freezer..

this is going to be epic


----------



## kcurnow

Looks pretty epic Jesse, to save material and cutting since it's a nice snug fit in your freezer you could reduce the depth of the long pieces so they are only half the height of the shorter cross pieces. This will reduce the amount of material required and cutting. If you were worried about stability you could only reduce the height of every second row or two.


----------



## Grainer

Froze some of my yeast yesterday.. FU&%ing keg fridge got too cold.. no glycerol... 10 strains down the toilet that were getting prepped for freezing.. fml..


----------



## Grainer

Yob said:


> so Ive decided to tackle storage, my current method is a bit random and it can be hard to find a vial of something in the 55lt container.
> 
> 
> 
> 
> DSC_1881.JPG
> 
> there is a fair bit of cutting involved but its going to be ******* stirling..
> 
> 15 x 11 and I can fit at least 2 layers in the tub which lives in the bottom of my 1100lt deep freezer..
> 
> this is going to be epic


Issue is .. I have those tubs and they crack really quickly in the freezer.. I am looking for new ones that will last the freezer ! but love the racking idea.. GOLD!


----------



## SBOB

Yob said:


> so Ive decided to tackle storage, my current method is a bit random and it can be hard to find a vial of something in the 55lt container.
> 
> 
> 
> 
> 
> DSC_1881.JPG
> 
> there is a fair bit of cutting involved but its going to be ******* stirling..
> 
> 15 x 11 and I can fit at least 2 layers in the tub which lives in the bottom of my 1100lt deep freezer..
> 
> this is going to be epic


im waiting for yeastdealzaustralia.com to open


----------



## Yob

None of mine have cracked and some are years old now.

In fact, I want to bring out a few vials from 2011 and see how they've held up.

YDA, had been considered


----------



## Yob

so in more of it's semi final glory..




with plenty of room left for the OG containers on top of that level

Im still toying with a couple of ideas to improve this concept


----------



## SBOB

Yob said:


> so in more of it's semi final glory..


come on... my inner OCD says you could at least co-ordinate cap lid colours in some kind of pattern


----------



## Yob

My inner OCD days I should be sorting by manufacturer and then ascending numbers


----------



## MartinOC

My inner OCD says you should also cross-reference the above & create a spreadsheet with your entire bank with freeze-dates & %age glycerine & be sure to save at least 3 copies of the spreadsheet in different places in the event of nuclear attack/bushfire/flood/cyber attack.

OK, maybe 4 copies (or 5 if you can get them all to update simultaneously, with a Master-copy held in a hermetically-sealed safety-deposit box under a rock, somewhere in Canada).


----------



## Yob

Ummm.. I have a spreadsheet...


----------



## MastersBrewery

MartinOC said:


> My inner OCD says you should also cross-reference the above & create a spreadsheet with your entire bank with freeze-dates & %age glycerine & be sure to save at least 3 copies of the spreadsheet in different places in the event of nuclear attack/bushfire/flood/cyber attack.
> 
> OK, maybe 4 copies (or 5 if you can get them all to update simultaneously, with a Master-copy held in a hermetically-sealed safety-deposit box under a rock, somewhere in Canada).


 surely a box burried 150km north east of Kal would do the trick.


----------



## Yob

Anybody have 50 or so 50ml vials they ain't using? 

I don't have enough to form my ice rack part 2 thingo...


----------



## MastersBrewery

Yob said:


> Anybody have 50 or so 50ml vials they ain't using?
> 
> I don't have enough to form my ice rack part 2 thingo...


 at first read I laughed so hard, then all I could think was here comes another BB, and fell off the couch laughing. Your your own nemesis.


----------



## Yob

missus said Im a yeast hoarder and I'll end up on a TV show one day...


----------



## mofox1

Yob said:


> Anybody have 50 or so 50ml vials they ain't using?
> 
> I don't have enough to form my ice rack part 2 thingo...


Lolz.

Aye, come grab mine. Would have 30 or so, at least. Mostly just use 15's now these days.

Dibs on YDA shares.


----------



## mofox1

Yob said:


> missus said Im a yeast hoarder and I'll end up on a TV show one day...


Slow on the uptake. I'll need when and what channel. Kids have commandeered 22.


----------



## Yob

Thanks Mick, it's club night tonight so might swing past on the way?


----------



## GrumpyPaul

Yob said:


> Anybody have 50 or so 50ml vials they ain't using?
> 
> I don't have enough to form my ice rack part 2 thingo...


You can have mine too if you want.

Think I've probably got closer to fiddy .


----------



## Yob

Woot!! Do you require a lift to club night?


----------



## GrumpyPaul

Yob said:


> Woot!! Do you require a lift to club night?


Yes please.

I shall pay my fare in yeast vials


----------



## mofox1

Yob said:


> Thanks Mick, it's club night tonight so might swing past on the way?


Done diddly.


----------



## Bruer

So a quick question. I'm getting ready to freeze some yeast later today. I'm just wondering about insulating the 50ml tubes in my frost free freezer. I've got some blocks of Styrofoam laying around from bbq packaging. If I drilled out some holes to fit the tubes snugly, do you guys think it would insulate properly against the defrost cycle? Or should I go the clip top tupperware thingo with foam lining and salt ice? Also is 35% glycerin the go for most people?


----------



## Yob

Bruer said:


> Or should I go the clip top tupperware thingo with foam lining and salt ice? Also is 35% glycerin the go for most people?


this... ^^^^^^

Im usually at about %25 now, just coz its easy numbers to do


----------



## Bruer

Yob said:


> this... ^^^^^^
> 
> Im usually at about %25 now, just coz its easy numbers to do


So that's 25% glycerin and 75% yeast slurry?


----------



## Bruer

How can I sterilise the glycerin without an autoclave/pressure cooker. I have plastic tubes.


----------



## Yob

Microwave


----------



## Yob

Bruer said:


> How can I sterilise the glycerin without an autoclave/pressure cooker. I have plastic tubes.


Wait.. Wait.. You have test tubes?


----------



## Bruer

Yob said:


> Wait.. Wait.. You have test tubes?


Plastic centrifuge tubes.


----------



## Yob

Perfek 

Microwave in a pyrex jug if you have one, half water and half glycerine... then 25ml in each tube.

Top up with yeast slurry, fridge 36 hours, freeze.


----------



## Bruer

Yob said:


> Perfek
> 
> Microwave in a pyrex jug if you have one, half water and half glycerine... then 25ml in each tube.
> 
> Top up with yeast slurry, fridge 36 hours, freeze.


Cool... I figured if it was good enough for doing with gelatin, it was good enough for the glycerine.  

And just soak the tubes in starsan before filling?


----------



## Yob

Bruer said:


> Cool... I figured if it was good enough for doing with gelatin, it was good enough for the glycerine.
> 
> And just soak the tubes in starsan before filling?


thats what I do mate, yep


----------



## Nullnvoid

How long in the microwave?


----------



## Yob

If it boils over... It's too long


----------



## Moad

I've just read this entire thread for the second time and am inspired to give it a go, gives me a good excuse to buy a pressure cooker for use on other brewing applications. 

Planning on pre making various strength starter wort jars and storing to make the starter process quicker and easier as well.

Do you guys typically keep a strain going for a few generations and then build up another frozen sample. Or do you capture each time? I can see if you captured each generation you could build up a huge bank quickly and not have to buy yeast for a long time.


----------



## Yob

I typically split and bank first generation into 5, then when I use it and it gets cracking, say third gen, I'll wash and save that as well.

When we talk generations, we say third but it's really about 18-20, each ferment will generate about 6-8 budding cycles.. But yeah.. Whateves.. We all know what we mean I think..


----------



## Moad

I'm following, just trying to work out how to use it effectively... bit of a case of RDW....

So first build a starter and split that out into 1st gen vials. Then for each vial build a starter and split that out again (25 vials of 3rd or 4th gen including steps). That should be enough for a strain for me as I will use slurry for a few brews (from 20-120L batches) So I could essentially get 100 batches out of 1 pack of yeast. incredible

Whats the longest you have had yeast stored and revived now? I believe my freezer is frost free, what is the process of the "salt water ice"? Literally making ice with salt water? I was thinking I could use zip lock bags with salt water and freeze them then pack them in insulated tupperware style container with the yeast.


----------



## Mardoo

You can go straight into vial from the manufacturer's package, or if you wish make a starter. From starters, I haven't seen the results of generational changes in the yeast during fermentation like I have seen it with fully fermented out yeast. For example a third generation yeast only grown in starters doesn't seem to show the same changes that a yeast fermented out to the third generation will. Does that make sense?

So you can save yeast from pack or starter and use that as first generation. After you've fermented it out three times freeze it as third generation. I've done that with 1469, which shows increased complexity in later generations, to my palate.


----------



## Moad

nice thanks Mardoo, makes sense.

So when starting up a vial I can capture some and refreeze and it wouldn't have as much drift as capturing some after fermenting out.

It is a whole hobby in itself!


----------



## Yob

it is.. and just as consuming as the parent hobby..


----------



## Mardoo

Yep, on the vial and the hobby. 

I've been wanting to geek hard and save out samples of 5 generations of the same yeast and ferment each generation on the same wort. Time permitting…HAH!


----------



## mofox1

Moad said:


> nice thanks Mardoo, makes sense.
> 
> So when starting up a vial I can capture some and refreeze and it wouldn't have as much drift as capturing some after fermenting out.
> 
> It is a whole hobby in itself!


That's what I do... build up starters to re-freeze while I'm building up to pitch into the next brew.

My process as follows:
- build up to a 2L starter (which is 3 steps in itself if coming from a single 15ml vial)
- crash chill
- decant
- pitch half into a new 2.5 / 3L starter (to pitch into wort)
- store the rest in vials for freezing

I prefer this method to yeast rinsing because the yeast is super fresh and (less important but more practical) I'm usually not organised enough to package beer AND capture yeast at the same time.


----------



## Yob

Im a little different..

depending on the age of the sample, and Ive still got a few from my first freeze that fire up with no issues, I'll do a 1lt starter (1.020) and then a 3lt starter (1.040), when that's going nuts, I'll pitch 2.5L to brew and tip 500ml into another flask and allow to settle out so that the yeast build their glycogen and trehalose reserves up, then split freeze.. and/or rinse and split if it was a particularly impressive ferment.

many many ways to beat that kitten to death.. try them all and work out what suits you best.


----------



## Moad

awesome exactly what I was after thankyou gents. 

Waiting on some vials to arrive and then I'll get my yeast on


----------



## Moad

Quick question, this seems to be the most relevant place to ask. I've got some Terumo disposable syringes from the chemist, they look to be older models of the Nipro which are apparently PP (autoclavable). I've seen some for 45 cents a pop but if I can reuse them...why not. Do you pressure cook the syringes and if so, how?


----------



## Yob

For transfer? I don't bother with syringes any more, just tip directly from flask or pack. Less process and equipment


----------



## Bruer

I love this thread - definitely catering to my inner nerd. I made up some extra wyeast American Ale II from a starter for my last beer. Froze them last week. Will have to wait and see if they'll fire up.


----------



## Yob

I pulled one out last night that's been in the freezer for 18 months, will note it's progress for folks over the weekend. I hope to have it on the plate tonight for a first step and into a 3L starter by tmoz night and pitch Sunday (+ restock 3 vials)


----------



## Yob

So this is is about 18 months in the deep freezer





with my 500ml flasks in the fridge with some 1728 in them I had to use a bigger flask for the first step of 1.020





at 12 hours there were clear signs of respiration which I thought was pretty good, the first step like this isnt really about building number but more restoring health.





so at the 12-18 hour mark I make a further 200g into 1.5L, chill and tip the whole lot in.





at 18 hours later I have this, Ive let it go till tonight and have turned the plate off.

I'll let this lot settle out of it's own accord before decanting and pitching toward the end of the week.

Overall, pretty happy with an 18 month old sample, Ive done older ones with very similar results and this is fairly typical of the reactions I get.

Cheers


----------



## Coldspace

That's so cool, 
Going to try this one day.


----------



## SBOB

Yob said:


> So this is is about 18 months in the deep freezer
> 
> 
> 
> Overall, pretty happy with an 18 month old sample, Ive done older ones with very similar results and this is fairly typical of the reactions I get.
> 
> Cheers


that semen sample really did take off


----------



## Yob

good swimmers, what can I say


----------



## VP Brewing

I found a box of these at work today. 500 of them! Autoclavible too.


----------



## Yob

win... keep 250 minimum.. 300 is a better number h34r:


----------



## VP Brewing

Yep I think 50ml is a better size than the 28ml ones I've been using which are a bit of a pain in the arse to revive. 
Growing up some 1272 as we speak. 
I've borrowed about 20 of them...


----------



## dblunn

VP Brewing said:


> I've borrowed about 20 of them...


Take the bloody lot mate, pilfering is a time honoured tradition


----------



## angus_grant

Damn it. Should not have read this thread. 
So time to buy some vials.....
Why are there so many expensive "money-saving" detours in the "cheap homebrew" rabbit warren?


----------



## Yob

"it'll pay for itself in no time"

The real advantage is just having all your favourite yeasts as a perpetual supply... The never ending list of equipment is of no consequence... 

Bulk glycerine 
Flasks of various sizes
Stirplates / yeast forge
Army of vials
Bulk dme (or frozen wort from ya brews)
Deep freezer space 

Whole other (worthwhile) hobby in its own right


----------



## angus_grant

I was lucky enough to be given some wy 1728 as I couldn't find any in aus. 
That's what triggered me into reading the thread.


----------



## Yob

Great yeast, worth banking for sure


----------



## angus_grant

I made a 2L starter and put 1.8L into my wee heavy, and let the rest settle out in a jar and put yeast into a vial. Got it sitting in the fridge. If I don't make another beer with it in the next month or 2, I'll run up a starter in a month or two and do the same.

Will need to read through the thread again and start purchasing supplies to start freezing yeast.


----------



## Yob

you can get a small amount of glycerine from coles, 200ml I think..

better to freeze as soon as you can to keep as much health in the yeast as possible, if you don't have any vials, shoot me your addie and I'll send you a couple up to get you started.

all you really need additional to what you probably already have is the glycerine and vials.


----------



## Yob

Yob said:


> So this is is about 18 months in the deep freezer
> 
> 
> 
> 
> 
> with my 500ml flasks in the fridge with some 1728 in them I had to use a bigger flask for the first step of 1.020
> 
> 
> 
> 
> 
> at 12 hours there were clear signs of respiration which I thought was pretty good, the first step like this isnt really about building number but more restoring health.
> 
> 
> 
> 
> 
> so at the 12-18 hour mark I make a further 200g into 1.5L, chill and tip the whole lot in.
> 
> 
> 
> 
> 
> at 18 hours later I have this, Ive let it go till tonight and have turned the plate off.
> 
> I'll let this lot settle out of it's own accord before decanting and pitching toward the end of the week.
> 
> Overall, pretty happy with an 18 month old sample, Ive done older ones with very similar results and this is fairly typical of the reactions I get.
> 
> Cheers


So as a follow up to this, I let it all settle out and pitched a cube last night, there was a little bit of yeast left in the flask so I thought I'd run out 1.5L

Overnight this happened





I know its a little mucky but I can just rinse the yeast at the end once it's dropped clear and re freeze.. looks like this 1217 will be going back into cryo in a few days


----------



## MartinOC

What temp. you running that Yeast Forge at? Difficult to see from the piccies & seems to vary from one to the next (unless I'm seeing spin-speed?). 

When CB gets a new shipment of those babies in, I'm grabbing one for myself  .


----------



## Yob

That was running at ambient, I've set it to 20'c now.. Thanks for the prompt


----------



## mattlea266

If you want one sooner they are here.
http://www.digitalhomebrew.com/yeast-forge


----------



## Curly79

mattyl said:


> If you want one sooner they are here.
> http://www.digitalhomebrew.com/yeast-forge


The bloke from Clever Brewing gives him a discount or something I think?


----------



## MartinOC

Curly79 said:


> The bloke from Clever Brewing gives him a discount or something I think?


----------



## Moad

Vials arrived and I got an old packet of wy1388 yeast to practice on.

I've got it on the stir plate now and the vials in the pressure cooker.

Do I need to sanitise the glycerine? Should I simply put the glycerine in the vials and then pressure cook them before adding the slurry? That would sanitise the vials and the glycerine


----------



## VP Brewing

Yeah mate. Glycerine and vials need to be sterilised. I fill them 1/4 with glycerin, 1/4 with water then sterilise. 15min at 100°C+ in a pressure cooker will do. 
Then they can be filled the rest of the way with yeast slurry and frozen. 
Hope that helps.


----------



## Moad

Yup perfect cheers


----------



## Yob

That's the spirit lads.. Into it


----------



## Moad

Successfully restarted some 3787 from last years tripel (possibly 2 years ago I cant remember).

Nice white layer of creamy yeast, now in the freezer with 25% glycerine, 25% water, 50% slurry to be spun up as a test in a month.

I put the vials in a container insulated with ice packs. I filled the container half way with salty water and can slide the vials in and out of their "slots".

Got some 1388 crashing to decant and then some Dennys Favourite that I'll start up for a brew and freeze a few vials. I think I'll get some ESB 1968 and some 3068. That should cover me for most of my brews and see me through a year or two.


----------



## Yob

It has begun... I take no responsibility....


----------



## Bruer

I've got ESB 1968 and American Ale II, just need a few more and I'll be mostly good too.


----------



## Bruer

Yob said:


> It has begun... I take no responsibility....


You're a bad man Yob.


----------



## Yob

Bruer said:


> just need a few more and I'll be mostly good too.


Yeah, that's what I said...


----------



## Moad

checked it this morning, the contents of the vial seem to be frozen... is that right?

Not solid, if I shake it up it becomes an icey slurry


----------



## Moad

I've just put some more yeast in the bank and noticed the original vials are frozen solid. 

It was 12-13ml out of 45-50ml. So 25-30% glycerine and it is still frozen.

Is that ok?

edit: nevermind I reread through the start of the thread and it looks like it will be fine.

Anyone in Newcastle interested in some Glycerine? I just purchased 5L for $9/L


----------



## angus_grant

You're a bad man yob. 
30 * 50ml microwavable tubes. 
Now a starter of wyeast Scottish ale and try spring some up. 
::slipperySlopeContinues::


----------



## Coldspace

Yes , slippery slope.....

I've got about 8 varieties in packs of 3 frozen down now. Handy when time or unavailability of a strain, can keep brewing. Most of my local shops only stock dried so this method has come in handy.

I just struck up to life a 1084 that's been down for 4 months now. No probs. I tend to run my glycerine at 35% to 40% total vial amount as the 50% slurry I add into the vial ontop of the glycerine water mix already has some water in it. Mine are at -18', and when I look at them they are not frozen up, just sort of in a really thick state. Just under a solid.

Tks to Bribie and Yob for paving the way.


----------



## Moad

Will be upping my glycerine mix to 40% from now on just to be safe


----------



## Yob

I went up to %50 and they remained fluid, however, it seemed at least anecdotally that ot took a lot longer on the plate to expel the glycerine first at start-up so I knocked it back to %25 and while they solidify, I do get seemimgly faster start times so have stuck with that ratio


----------



## Mardoo

In the yeast book Chris White and Jamil Z make the point that the problem with freezing yeast is dehydration of the yeast cells. The glycerine keeps them from dehydrating, not freezing. If your vial freezes solid it isn't a problem, so long as the yeast are stored in a glycerine solution. Mine freeze solid at 25%, and kick off just fine. I too have noted the greater lag at around 50% glycerine.


----------



## Coldspace

I'll keep mine at approx 30% glycerine then. 20% sterile water, 50% thick yeast slurry.

I use 30 ml vials I got from my local chemist for 40 cents each. After I fill em up and lid em, I wrap the lid around with white electrical tape. This seals the lid tight and gives me something to wright on instead of the lid, then I can re- use them.

I found also after filling these up half way with boiling glycerine/water mix, I lid em and drop them straight into a dish of water to cool quickly , otherwise I have noticed the plastic vials can warp slightly and also cools it quicker to then get the slurry ontop faster.

Many diff ways to skin a cat.....

My 1084 struck up good at the 40% mark but it was only 4 months old, as this is when I first put them in. Got some nice British ale yeasts coming in, will propagate them up, then freeze em at 30% mix.

Awesome......


----------



## Moad

OK 25-30% it is then.

I also just took delivery of 5KG of glycerine. Anyone need some? Can ship 1KG for $25 (glycerine + shipping).

Or pickup for $10 a KG in Newcastle


----------



## MastersBrewery

Finally got me a pressure cooker now just need to track down some vials and give this a red hot go. 
Do any of you guys wash yeast from a 1st gen ferment and split? Or is it safer to just split from fresh packs?


----------



## Yob

Both.. Depends on my timing and the ferment


----------



## MastersBrewery

Yob said:


> Both.. Depends on my timing and the ferment


 Ahhuh clear as RIS.


----------



## SBOB

Moad said:


> Or pickup for $10 a KG in Newcastle


I'll grab 1 L/kg off you.. Im local (Wallsend) but away for work this coming week but can organise to pickup at some stage?



Anyone got a recommendation/source for cheap vials that are pressure cooker/microwave safe?


----------



## Yob

Check the bulk buys, Martin oc ran one a while back and I've only got praise for the vials, they rock


----------



## dr K

Is it really worth the effort?
Last millennium when I started brewing liquid yeast was hard to get, often old and bloody expensive.
These days there are many local suppliers who stock fresh liquid yeasts, some at $9 or less a pack.
80 billion or so viable cells fresh from the lab....mmmmmm

K


----------



## Yob

Not as fresh as right off the plate, I find having a library invaluable, I don't have to order in advance (sure, I've gotta stir it up though) and I've got an eternal supply of my favourites,many of which are PC strains and aren't available all year round, some aren't even available in Australia.. 

So yes... Totes


----------



## Ducatiboy stu

dr K said:


> Is it really worth the effort?
> Last millennium when I started brewing liquid yeast was hard to get, often old and bloody expensive.
> These days there are many local suppliers who stock fresh liquid yeasts, some at $9 or less a pack.
> 80 billion or so viable cells fresh from the lab....mmmmmm
> 
> K


I remember those days...

And 10yrs ago a wYeast smack pack was about $16....if they had them

But in saying that, this whole freezing yeast thread has been very interesting

And the idea of having a library in the freezer does have some merits, especially if you only want to use a specific yeast say once a year for a seasonal brew


----------



## MastersBrewery

dr K said:


> Is it really worth the effort?
> Last millennium when I started brewing liquid yeast was hard to get, often old and bloody expensive.
> These days there are many local suppliers who stock fresh liquid yeasts, some at $9 or less a pack.
> 80 billion or so viable cells fresh from the lab....mmmmmm
> 
> K


when you live more than 100km out side a capital city the only choices in yeast are dried and very limited in range. I do go into the city every few months so when I do I'll be grabing a few strains to put away. Most everything else I get in bulk and therefore have on hand. Which reminds me I need some 3068 before spring.


----------



## Ducatiboy stu

100km...fark...that is a bit close for my liking


----------



## SBOB

just grabbed a bunch of vials from china (1/2 the price of proscitech), so in a month or so I'll be joining the yeast freezing bandwagon (after I hit up Moad for some glycerine)


----------



## gezzanet

Got a link for those vials sbob?


----------



## Mardoo

Been thinking its time for a ProSciTech bulk buy…let me look into it.


----------



## SBOB

gezzanet said:


> Got a link for those vials sbob?


http://www.ebay.com.au/itm/351748453646

All the others are similar price to proscitech, but this seller charged no extra shipping on top of the first 10...
so $20 for 60x50mL vials


----------



## SBOB

so, how are people storing their bulk glycerine supplies (assuming you have it not in the container it was shipped in)?

Good enough to just store in sterilised containers, or would go through some kind of pressure cooker/autoclave process to sterilise the glycerine also?


----------



## Yob

I've got it in a keg king growler 

Just a PET bottle would do


----------



## MartinOC

Keep it in the container it arrived-in.

Mix-up your 50/50 in a pyrex jug & microwave it, then divvie it up into your pre-sanitised vials.

Once cool, add your yeasties & freeze.


----------



## Moad

It gets sanitised if you pressure cook it in the vials with your mix so what it came in should be fine sbob


----------



## SBOB

SBOB said:


> http://www.ebay.com.au/itm/351748453646
> All the others are similar price to proscitech, but this seller charged no extra shipping on top of the first 10...
> so $20 for 60x50mL vials


Arrived today, so about a 2 week shipping time. 
Don't have any other to judge quality against, but the look like vials/tubes to me. 

https://goo.gl/photos/TUgqZV4aBp87ZL157


----------



## Yob

Get yourself some round stickers for the lids so you can read them easily..

Thems the shiz


----------



## fishingbrad

SBOB said:


> Arrived today, so about a 2 week shipping time.
> Don't have any other to judge quality against, but the look like vials/tubes to me.
> 
> https://goo.gl/photos/TUgqZV4aBp87ZL157


Cheers mate. I ordered mine a week ago, so another week to go. Got some fresh 1084 & 1272 just itching to be frozen.


----------



## SBOB

Yob said:


> Get yourself some round stickers for the lids so you can read them easily..
> Thems the shiz


Confucius say
"Man who freeze yeast, listen to Yob advice.."

Will do...


----------



## SBOB

so, whats the current dummies guide in step by step process?

- combine glycol + water at a 50/50 ratio
- fill 50ml vials ~1/2 way (making it 10ml glycol, 10ml water)
- sterelise vials (e.g. Pressure cooker for ~15 mins), let cool

Meanwhile (assuming i have some yeast currently sitting in jars in the fridge)
-create basic yeast starter to wake them up, build up numbers etc (any ideas on yeast size starter?)
- wait till fully fermented out, then chuck in the fridge for a day or so to settle
- decant off excess wort/beer
- top up vials with ~20ml of yeast slurry

Vials now full with 25% glycol, 25% water, 50% yeast
Into the fridge for ~2 days, then into the freezer


Have I missed anything? Anything I should do tomorrow?


----------



## Yob

Sounds about right to me mate.. Get amongst it


----------



## MartinOC

SBOB said:


> so, whats the current dummies guide in step by step process?
> 
> - combine glycol + water at a 50/50 ratio
> - fill 50ml vials ~1/2 way (making it 10ml glycol, 10ml water)
> - sterelise vials (e.g. Pressure cooker for ~15 mins), let cool
> 
> Meanwhile (assuming i have some yeast currently sitting in jars in the fridge)
> -create basic yeast starter to wake them up, build up numbers etc (any ideas on yeast size starter?)
> - wait till fully fermented out, then chuck in the fridge for a day or so to settle
> - decant off excess wort/beer
> - top up vials with ~20ml of yeast slurry
> 
> Vials now full with 25% glycol, 25% water, 50% yeast
> Into the fridge for ~2 days, then into the freezer
> 
> 
> Have I missed anything? Anything I should do tomorrow?


You forgot to relax, have a beer & be happy in the knowledge that you've done everything right.

Tomorrow, have a beer & be happy that you've done everything right.


----------



## MastersBrewery

Getting the last bits in to kit off my little bank. As above vials are in the post.

Was wondering if anyone has split and recultured Wyeast 3068 successfully?


----------



## SBOB

MartinOC said:


> You forgot to relax, have a beer & be happy in the knowledge that you've done everything right.
> 
> Tomorrow, have a beer & be happy that you've done everything right.


I do that every day after work already


----------



## MartinOC

SBOB said:


> I do that every day after work already


Ditto...


----------



## Alex.Tas

MastersBrewery said:


> Getting the last bits in to kit off my little bank. As above vials are in the post.
> 
> Was wondering if anyone has split and recultured Wyeast 3068 successfully?


I've split and stored it in the fridge for around 8 months and restarted it without any worries. I don't have space in my freezer, so can't comment on reviving a frozen sample.


----------



## Exile

Anyone know how would plastic bottle blanks go for yeast freezing?


----------



## Yob

great I suspect, they wont stand up by themselves though


----------



## Exile

Yeah that's the only problem I guess.
I got given a garbage bag full of them today, and the guy said he has trouble getting rid of wool bales full of them, as the water bottle machinery is made in china and stuffs up all the time.


----------



## Ducatiboy stu

SBOB said:


> so, whats the current dummies guide in step by step process?
> 
> - combine glycol + water at a 50/50 ratio
> - fill 50ml vials ~1/2 way (making it 10ml glycol, 10ml water)
> - sterelise vials (e.g. Pressure cooker for ~15 mins), let cool
> 
> Meanwhile (assuming i have some yeast currently sitting in jars in the fridge)
> -create basic yeast starter to wake them up, build up numbers etc (any ideas on yeast size starter?)
> - wait till fully fermented out, then chuck in the fridge for a day or so to settle
> - decant off excess wort/beer
> - top up vials with ~20ml of yeast slurry
> 
> Vials now full with 25% glycol, 25% water, 50% yeast
> Into the fridge for ~2 days, then into the freezer
> 
> 
> Have I missed anything? Anything I should do tomorrow?


Now if that was instructions on how to make good beer it would be 4 pages long, what with all the micro steps that everyone would want included.


----------



## Yob

Exile said:


> Yeah that's the only problem I guess.
> I got given a garbage bag full of them today, and the guy said he has trouble getting rid of wool bales full of them, as the water bottle machinery is made in china and stuffs up all the time.
> 
> 
> 
> 
> 
> blanks.png


I'll take a shopping bag


----------



## Ducatiboy stu

Exile said:


> Yeah that's the only problem I guess.
> I got given a garbage bag full of them today, and the guy said he has trouble getting rid of wool bales full of them, as the water bottle machinery is made in china and stuffs up all the time.
> 
> 
> 
> 
> 
> blanks.png


If you want to off load a few I might get into freezing some yeast.....Note to self...buy some w1728


----------



## Mardoo

They most likely won't stand up to pressure cooker temps, if you use one.


----------



## Ducatiboy stu

Mardoo said:


> They most likely won't stand up to pressure cooker temps, if you use one.


Mmmm...they might


----------



## Yob

The old white labs vials did


----------



## Camo6

Yob said:


> The old white labs vials did


Never tried it myself but read they tend to get a bit soft and distort? Then again, so did my autoclavable type when I left the lid tight... :unsure:


----------



## SBOB

so, first attempt at creating some vials of yeast to freeze. Currently sitting in the fridge (or yeast transfer station for Yob)

Do these look like viable samples. Seems a little low on the 'solids' part compared (was 40ml of water/glycol and 40ml of slurry from a starter).. Perhaps too watery a slurry?

https://goo.gl/photos/UqqssHsgUySdKm547

Its a Wyeast 1056 batch, so not the end of the world if its too small a sample.. next attempt is a Wyeast 1217 (West Coast IPA, which is a limited release), so hoping to identify any issues on this round


----------



## Mardoo

They'll be fine. You can start from a speck. I myself put a bit more than that.


----------



## SBOB

Mardoo said:


> They'll be fine. You can start from a speck. I myself put a bit more than that.


if you are decanting from a starter, how do you manage to get a larger sample/thicker slurry?


----------



## Mardoo

Into a 50ml tube I put 25 ml of 50/50 water/glycerine solution. Pressure cook. Decant spent wort from starter leaving either a bit of wort, or fully decant spent wort and use sterile water to thin slurry as you wish. Add 25ml of the slurry. My slurry usually ends up around 50% liquid, 50% yeast, so you end up with 12.5ml of yeast, or so. It's not precise.


----------



## Moad

Fired up my first frozen tube today. Pulled it out of freezer Wednesday then out of the fridge today, let it warm to room temp then into 200ml of 1.020 wort. On the stirplate now. Next time will likely start with 50ml then into 200.


----------



## SBOB

I did one this week.. Sample only frozen for a week though, but 500ml 1.020 and then 2l 1.040 steps kicked it off well...


----------



## Yob

Moad said:


> Fired up my first frozen tube today. Pulled it out of freezer Wednesday then out of the fridge today, let it warm to room temp then into 200ml of 1.020 wort. On the stirplate now. Next time will likely start with 50ml then into 200.


what volume of yeast solid?


----------



## Moad

About 5ml in the tube when it dropped out


----------



## Moad

Its alive!!! Only frozen for 2 months but there is a nice layer of yeast after 2 days, just stepped up to 1.8 L at ~1.040. 

Will then pour some off to be rinsed and refrozen and pitch the rest. Awesome


----------



## fishingbrad

I've got Vials prepped as per above however I did not shake the vials after refridging for 24hrs, and now frozen I can see the yeast has separated from the mix. Is this normal ? or should the yeast be mixed in solution ?

Cheers.


----------



## Yob

its normal for them to floc out,

RDWAFTV


----------



## Yob

The Yeast Bank - Coming soon to a freezer near me h34r:


----------



## SBOB

patent pending


----------



## Mardoo

Yob said:


> The Yeast Bank - Coming soon to a freezer near me h34r:


My freezer is near your place


----------



## Yob

Ha.. I'll need a prototype tester


----------



## Moad

damn this thread, 40 more vials, a yeast forge and a freezer are on their way.

It's real nice having a choice of yeasts only a few days away whenever I like.

cheers to all who have contributed!


----------



## Yob

Make beer for cheap they said...

You'll save money they said...

You don't NEED much equipment they said...


----------



## MastersBrewery

Yob said:


> Make beer for cheap they said...
> 
> You'll save money they said...
> 
> You don't NEED much equipment they said...


----------



## Camo6

Spun up a couple of vials from the freezer the other day. One was top-cropped 1469 and the other some recultured Greenbelt from the Yobster. Both were dated around Jan 2015. Stepped from 50mls to 500mls and they were climbing out of the flask within a couple of days. That Greenbelt is a monster.


----------



## Coldspace

It's easy as isn't . I love my yeast collection now. Got some more samples to grow up and freeze. Isn't really about cost, just awesome to have on hand a frozen selection for diff brews. Great for busy lifestyle, can start up a started, and pitch 48 hrs later . Especially when most of my local shops only carry dried stuff.
Thanks again for those who led the way on this new hobby/ obssesion lol

Only problem I've got now, is fighting the missus to keep a shelf in our freezer free for me lol


----------



## MastersBrewery

First world issues..... 
I can get 20kg of dme locally for $45. But how the hell do I store it. Do I vac seal down to 2kg lots?


----------



## AJS2154

20kg for $45......isn't that really cheap? I would get some at that rate and have a crack. Why would it not keep if vacuum sealed, it is away from pests, humidity and sunlight.


----------



## SBOB

MastersBrewery said:


> First world issues.....
> I can get 20kg of dme locally for $45. But how the hell do I store it. Do I vac seal down to 2kg lots?


thats crazy cheap
from where?


----------



## MastersBrewery

Yarragon, Latrobe Valley.
That a lot of vac sealing, it's not like I'm some exotic hop merchant. I'm moving house next week so I'll grab some but wait till the boss's are happily settled.

Ed, getting used to reading glasses missed a post.


----------



## mofox1

Vac seal not really necessary... Can just bag up into the biggest ziplocks you can find. I've stored approx 10kg in a bucket for the past year.. almost empty from yeast starters and OG fuckups. Likewise for the previous year. It's fine until the last few cm, then it was noticeable it has started to pick up a bit of moisture from the air.

Just about to finish this bucket and I've already got the dme to fill it again, not going to do any different this time.

But $45.. jeez.


----------



## MastersBrewery

Now you guys have me worried; I didn't have the new reading glasses when I saw that price!?!??


----------



## mofox1

At that price I'd be checking it actually is malt extract, and not just dex/glucose. Keg king price for 25kg of dex is $50, vs $120 something for DME. Not necessarily the best prices, but won't be far from it.


----------



## Yob

I've got to pick up a new bag too, I've gone through 20kg just in starters over the last year or so.. I just kept it in the bag with a big moisture absorber pack on top, still dusty to the last. 

Under $50 is a steal, not sure I'm even getting it for that


----------



## VP Brewing

Yob said:


> So this is is about 18 months in the deep freezer
> 
> 
> 
> 
> 
> with my 500ml flasks in the fridge with some 1728 in them I had to use a bigger flask for the first step of 1.020
> 
> 
> 
> 
> 
> at 12 hours there were clear signs of respiration which I thought was pretty good, the first step like this isnt really about building number but more restoring health.
> 
> 
> 
> 
> 
> so at the 12-18 hour mark I make a further 200g into 1.5L, chill and tip the whole lot in.
> 
> 
> 
> 
> 
> at 18 hours later I have this, Ive let it go till tonight and have turned the plate off.
> 
> I'll let this lot settle out of it's own accord before decanting and pitching toward the end of the week.


I've just finished these steps, starting Monday night using 1275. Planning on fermenting the vic swap brew with it then using the cake for a RIS . 
Pitched the 500ml @ 1.020 flask into 1500ml @ 1.040 the next morning (12 hrs) after seeing the signs of respiration on the sign of the flask. It took almost 72 hours to get any krauzen. Checked this morning. 
Would this lag time be stressful to yeast health in any way? I think next time I will let the first step go a bit longer.


----------



## Yob

each culture on its own merits mate, I tranfered only when I was sure it was ready, times vary.. I was going to throw one out a few back, was 48hrs with little signs... then boom! away it went.

If they are excessively slow (or even a bit longer than I expect) I'll let the second starter finish the ferment, as in, once Ive seen growth and a krausen, I'll switch off the plate and let them ferment and settle.

This allows all the yeast to build up their reserves and gets them to optimal health.

Then pitching to another 2lt starter has always shown expected results within hours. Its a method of 'proving' the yeast and Ive not had a fail yet with this method. But in saying that, I rekon at times they yeast have needed to be 'restored' with care.. one of the problems I have is that early on I was testing various amounts of glycerine concentrations and Ive no fekin idea which is which and as the concentrations got higher it took longer for the yeast to dispel it and get back to health and full operation.

Its the bed Ive made for myself so Im aware that each vial may react differently so I approach it fairly pragmatically and not in a hurry


----------



## Yob

VP Brewing said:


> Would this lag time be stressful to yeast health in any way? I think next time I will let the first step go a bit longer.


I dont believe so, you dont want it in your beer but a starter is a different ball game, particularly if you are decanting and pitching slurry.


Do make sure your adding a bit of yeast nutrient to each starter too, that is always a good thing to be doing.


----------



## VP Brewing

Cheers Yob. Great info. I've just got home an taken it off the stir plate. Will get another 2L ready to pitch into with nutrient this time. Couldn't be arsed going out to the shed to get it for the others and didn't think it mattered.


----------



## Yob

its still a pretty harsh environment for them so a bit of love here and there can go a long way


----------



## MastersBrewery

Time love and tenderness. Yob you know the song....


----------



## Killer Brew

I may need to look into freezing. I collected a nice active cup of weinstephen yeast from a wheat beer I brewed around 4 months back. Rinsed multiple times until clean and stored it refrigerated and airtight. Tried to get it going in a 1L starter at 1036 last week but no good. Seems it was all dead so 4 months is obviously a stretch in the fridge!


----------



## MartinOC

Yob said:


> its still a pretty harsh environment for them so a bit of love here and there can go a long way


In a word - Yeast-Forge is your friend (OK, OK, it's hyphenated & technically not a single word, but you guys get the gist, right?). :super:


----------



## Yob

MartinOC said:


> In a word - Yeast-Forge is your friend (OK, OK, it's hyphenated & technically not a single word, but you guys get the grist, right?). :super:


FTFY


----------



## MartinOC

No, you silly person! Go & change your armour!

Gist: :the substance or general meaning of a speech or text."

There's an extra "r" in there that you added that might have escaped your ken....Go & have another glass of RIS ya wazzock!


----------



## Yob

Insert facepalm....


----------



## Curly79

I'm in! Wish me luck. Freezer says its sitting at -18c. Is that a good temp or should I crank it up a bit?


----------



## Yob

that's good man, they'll pretty much sleep below 0'c


----------



## Curly79

Sweet. Got my little vials. That fella who lives round the corner who know a thing or two about home brewing has given me a few pointers. Gunna get a 1.040 starter going with some WL Burton Ale Yeast. How many vials do you think I need to prepare for a 
1Ltr Starter?


----------



## Yob

5-6.. you wont get much growth from 1L

that will equal 20ml in each of yeast/slurry roughly if you get the thickness right.. (like slightly warmed honey)


----------



## Curly79

Should I decant then re-pitch from the original starter into a fresh one or is 5 vials all I can expect from one sachet of yeast?


----------



## Yob

If the sachet isn't old, I'd just split into 5, freeze 4 and make a starter from 1..

You then have 4 pure pitches which you'll do starters on anyway..

Ya dig?


----------



## Curly79

Righto[emoji106]. Thanks again mate.


----------



## Bruer

Hey all, I've just made a batch of black IPA. The plan is to split it (22L/2) and pitch US-05 and wyeast B. lambicus. Has anyone had success freezing brett? Given the small batch size I figure I won't need to pitch all the brett, so taking 50ml should be fine I guess.


----------



## Yob

Dunno, think it was asked by those QLDers back a ways, not sure what came of it.


----------



## Yob

Version 2 point something..

5 x 5 to suit 50ml vials, stack-able pinlock on top so air can get under the one on top..

I can just fit in the 3D printer at work.. if I can get away with it..


----------



## Exile

Followed your steps Yob
Had about 2 ml of left over yeast and decided to just mix it in with the last vial I had prepared with the 50 - 50 glycerine & water mix. That was 2 weeks ago and checked it last Friday and noticed the vial looked like it was not quite frozen.
Put it in 600ml of dme for 12 hours and then decanted that into 1 liter of dme yesterday afternoon..
Just checked on it...... its alive


----------



## Yob

welcome to the club!

:super:


----------



## Exile

Forgot to mention its WLP008 East Coast Ale Yeast


----------



## Mardoo

Yep, if you do the 50/50 glycerine water it often won't freeze.


----------



## Yob

But will take longer to expel the glycerine from the cell wall


----------



## eldertaco

Has anyone managed to find glycerine in bottles bigger than 100ml? I've tried a few chemists but they all seem to only stock 100ml bottles of the stuff.


----------



## Yob

Mardoo did a bulk buy a little while back, maybe touch base there


----------



## sp0rk

I'll ask around at work if we know any good sources


----------



## Mardoo

This is the best price I found locally, at the 20L level. Brings it to $12.10 a litre. I haven't looked since though. There was a mob in QLD with a better price, but they never filled my order and stopped responding to emails, so...


----------



## eldertaco

Nice one Mardoo, I found that place earlier but I think I just found cheaper here.

edit: they have a bunch of interesting co2 extracts too, anyone for siberian fir winter ale?


----------



## SBOB

Moad bought some recently, as I bought some off him... So maybe he can let you know where he get some from


----------



## Lord Raja Goomba I

Any in Qld?


----------



## Coldspace

200 ml ones from Coles


----------



## Mattrox

Take 2 at my effort to reculture some coopers yeast and reculture in order to freeze some. 

I am methodically collecting yeast.

I have 500ml of 1.048 wort left over from previous brew. Will dilute with cooled boiled water to 1.020 to pitch a 6 pack worth of yeast.

I'm away on a work junket to Falls Creek on the weekend so reculture next week for brew day in 8 days time.


And hopefully 2 vials to freeze.


----------



## peteru

If you switch to drinking red Coopers, not only will you get more flavour, but you'll also get more yeast per bottle. :icon_cheers:


----------



## Moad

Yup I picked up the 5KG tub from sydney essential oil co linked above


----------



## Mardoo

That's a great price at the Sydney place!


----------



## lael

Yob said:


> Version 2 point something..
> 
> 5 x 5 to suit 50ml vials, stack-able pinlock on top so air can get under the one on top..
> 
> I can just fit in the 3D printer at work.. if I can get away with it..


Is there a sistema cliplock plastic box that fits the 50ml vials yob? I'm interested in making an insert to keep them all upright...


----------



## malt junkie

Yob is building a kit, Linky
To share/sell.


----------



## Yob

I powered up a 3yr old 1728 yesterday 500ml (1.020) 3000ml (1.040) and is going gang busters at less than 48hrs..

3 ******* YEARS!!

Anybody got the ability to lab test some of this output? very interested to test..


----------



## SBOB

Yob said:


> I powered up a 3yr old 1728 yesterday 500ml (1.020) 3000ml (1.040) and is going gang busters at less than 48hrs..
> 
> 3 ******* YEARS!!
> 
> Anybody got the ability to lab test some of this output? very interested to test..


you'll be bringing Walt Disney back to life soon enough


----------



## Yob

SBOB said:


> you'll be bringing Walt Disney back to life soon enough


edit: almost 3 years


----------



## malt junkie

Martin got me some fresh 001 and 051out to me arrived today all goes well I'll be splitting and spinning by lunch tommoz.
Thanks to all for the great info.
out here I'm lucky to find US05 in the closest brew shop!

Yob once they get to 4 you really should buy birthday present for them.


----------



## Midnight Brew

Yob said:


> I powered up a 3yr old 1728 yesterday 500ml (1.020) 3000ml (1.040) and is going gang busters at less than 48hrs..
> 
> 3 ******* YEARS!!
> 
> Anybody got the ability to lab test some of this output? very interested to test..


3 ******* YEARS, you know you want to throw it at a RIS.

I've got some Wy1318 coming up to 3 years in January. RIS bound.


----------



## SBOB

anyone frozen a dried yeast strain?
Currently doing a lager with w34/70 and was thinking I could get some of the yeast cake and putting it in the 'bank'

If so, any tips as Ive only done it from a fresh pack or a starter currently


----------



## Yob

Sure, why not, just make sure to rinse it well


----------



## gezzanet

Was going to do the same with 34/70 by letting it settle out from the yeast cake slurry first. This has been hibernating in my fridge for 4 weeks now. I might pour off the beer and freeze some to see how it goes. Colour is from a Speights golden ale clone for a kiwi mate. So hasn't gone dark due to age.


----------



## SBOB

Yob said:


> Sure, why not, just make sure to rinse it well


is there a dummies step by step on doing that?


----------



## LiquidGold

There sure is thanks to Wolfy.

http://aussiehomebrewer.com/topic/55409-rinsing-yeast-in-pictures/


----------



## Yob

SBOB said:


> is there a dummies step by step on doing that?


If you have a 2l/3l flask, put in 150-200ml slurry, top up with Sanitised water and shake/swirl vigorously, wait 20mins and pour off the top 3/4 into another flask/container. 

Rinsed 

Idea being all the trub and dead yeast will largely settle out in that 20mins. Most folks make the mistake of putting in too much slurry and making it too thick to settle easily.


----------



## malt junkie

After loading another 6 vials into the freezer today, I was skimming this blokes ebay store and saw this and though of this thread. Just in case your freezer ain't cold enough!


----------



## Mattrox

Yob said:


> Sure, why not, just make sure to rinse it well


Rinsed Coopers bottle yeast taken from the FV after my pale was bottled.

Rinsed enough?


----------



## peteru

That looks great. Now let it compact in the fridge to get better density. A smaller container will be easier to store.


----------



## Mattrox

peteru said:


> That looks great. Now let it compact in the fridge to get better density. A smaller container will be easier to store.


I'm going to step this up to pitch into my next brew, but save an aliquot to freeze


----------



## Mattrox

Ok. So the mason jar of yeast got a nice compact layer on the bottom. I poured off the liquid, then resuspended the yeast with cooled boiled water.

I used a glass sample syringe to draw up of the slurry and saved some in a vile. It looks like 15mL of uncompacted yeast.

I pitched the rest into a starter on the stir plate. Yes, there is a nice vortex under that krausen. It kicked off quickly.

Thinking I'll do a sneaky brewday tomorrow to pitch active yeast.

I'll do the glycerine shortly and freeze. 

Next question will be, what size starter should I pitch 15mL into after thaw?


----------



## Yob

500ml > 2000ml depending on age


----------



## Mattrox

Yob said:


> 500ml > 2000ml depending on age


1.020 for 500ml then step up to 2L @ 1.040?

Sorry about all the newbee questions. Another aspect of homebrew to get my head around. But rewarding.


----------



## Yob

Mattrox said:


> 1.020 for 500ml then step up to 2L @ 1.040?


that will sort you out nicely


----------



## Mattrox

Using *almost* sterile heating and *kitchen* aseptic techniques I prepared some vials of 50% glycerine.

I'm going to try to freeze some dry yeasts saved after fermentation as guinea pigs.

And I'll have a preserving jar of glycerine mix saved for later.


----------



## Mattrox

Ready to make a deposit in the yeast bank.


----------



## Yob

Why only half full?


----------



## Mattrox

Yob said:


> Why only half full?


Had yeast in fridge prior. It compacted so I poured off old water and put glycerine right on top.

Would have been been 40+ mL if yeast were collected straight from washing.


----------



## lael

I've been looking for a box to store the 50ml vials in for a while. Just found the 3L 'Really useful Box'es at Masters are the perfect height both for the vials and my freezer drawers. Hold appx 23 vials with 3 sitting up a little. $10 with 30% off = $7per box.


----------



## Moad

Yob when do you expect the yeast boxes to go into production? Might be worth holding out Lael they are very convenient...


----------



## Moad

48 hours in 250ml @ 1.040 (I already had it made up but normally do 1.020) then this after <24 hours in 1.8L @ 1.040.




Washed yeast that's been frozen for 6 months.


----------



## Yob

lael said:


> I've been looking for a box to store the 50ml vials in for a while. Just found the 3L 'Really useful Box'es at Masters are the perfect height both for the vials and my freezer drawers. Hold appx 23 vials with 3 sitting up a little. $10 with 30% off = $7per box.


Linky?


----------



## Yob

Moad said:


> Yob when do you expect the yeast boxes to go into production? Might be worth holding out Lael they are very convenient...


At $7 I'd get some of those too if they're any good, 

Production? Havnt settled on a bloody material yet, there are a few options... Don't get me started


----------



## lael

No link. Masters are in a fire sale. There weren't any left up here when I last went in. They are the small size of the 'really useful box'. I got two and wish I got more.


----------



## lael

Wait.. Did some searching. This is the box!: http://www.officeworks.com.au/shop/officeworks/p/really-useful-3l-box-clear-mfb3c#!specifications


----------



## Mardoo

Winter is coming…


----------



## SBOB

Wyeast changed their packaging?


----------



## Mardoo

And their website.

Edit: Just to clarify, the substance and structure of the packaging seems the same. It's not a major change like White Labs


----------



## Midnight Brew

Wyeast 1450 frozen on the 9.11.2014 with a 15% glycerine solution.

15ml vial > 100ml by 1020 starter > 1.2L by 1040 starter.

2+ years!!!


----------



## kcurnow

Midnight Brew said:


> Wyeast 1450 frozen on the 9.11.2014 with a 15% glycerine solution.
> 
> 15ml vial > 100ml by 1020 starter > 1.2L by 1040 starter.
> 
> 2+ years!!!
> 
> 
> 
> 
> 
> IMG_0422.JPG


2 years!!! Thats ridiculous, I may have to defrost some of my longuns and see if there is any life in them also.


----------



## Midnight Brew

Got some 1318 that I'll fire up in the new year for a round of milds. By that point they will be at the 3 year mark. 

Zombie apocalypse come at me.


----------



## Lethaldog

Can someone please explain the process to me in simple form cos .... well I'm a little simple[emoji23] I'd like to give it a go as I know how to grow and split smack packs but I don't brew enough to not consider freezing!

Cheers
Leigh


----------



## Mardoo

1. Get 50ml autoclavable vials from a lab supply place.
2. Add 25ml of 50/50 glycerine/water solution.
3. Put caps on loosely
4. Pressure cook for 15 or so minutes
5. Let cool
6. Get smack pack
7. Smack pack
8. Let swell
9. Get 5 of your 50ml vials of solution per smack pack
10. Pour contents of pack evenly into vials
11. Let vials rest 2 days in fridge
12. Shake vials well
13. Put vials into a container filled with ice and allow to freeze. It may not go solid because of the glycerine
14. Thaw when you need to use and pitch into 1.020 wort for first step, 1.040 for subsequent steps.
15. Grow yeast

There may be a stirplate involved in that last bit. Ideally, but not absolutely required. I think that covers it.


----------



## Lethaldog

Awesome Mardoo I just screen shotted that shit for a later date cos I have one eye closed at the moment just trying to focus ( not quite but close) thanks I will definitely be having a go at that!

Pressure cook for 15 minutes? Do I need to get a pressure cooker or just boil in a pot?

Put vials into a container filled with ice? 
Is that strict or can I just put them in the freezer?

Does it matter if the vials are plastic or should they be glass?


----------



## angus_grant

You can get plastic vials which are auto-clavable pretty cheap. 
I've been microwaving my vials as I don't have a pressure cooker. Same process as Mardoo listed though.


----------



## Lethaldog

angus_grant said:


> You can get plastic vials which are auto-clavable pretty cheap.
> I've been microwaving my vials as I don't have a pressure cooker. Same process as Mardoo listed though.


how long would you need to microwave them for?


----------



## Mardoo

There are a fair few "good practice" things in yeast freezing you can probably sneak past if you want, but I figure they're simple enough that the added effort is minimal.

Pressure cooking pretty much guarantees no possibility of infection from encysted or spoored bacteria. Boiling is probably good enough, but I'd definitely go Angus' microwave method over boiling, once he tells you what it is. I'm not willing to risk probably, myself. You can pick up a pressure cooker for $10 or so from an op shop. I got mine for $12. Aldi sells a cheap one that lots of folks on this thread use.

You don't have to put the vials into a container with ice when freezing. However, the faster the yeast gets to freezing temperatures, the better the viability you should get. Pretty sure that was from the yeast book, but I may have seen it in a study. Can't remember. It's not uncommon for a vial to not actually freeze if it's ended up with an extra bit of glycerine. However, it's at freezing, and I haven't yet had problems with viability from one of these vials.

You don't have to use glass vials. You can if you can find ones with secure lids.


----------



## angus_grant

I've only done a few batches so not really commenting from experience. Haven't actually roused any yeasts from frozen sleep yet, so grain of salt....
Microwaved for 10 mins. Not melted any vials yet. 
Pretty sure yob (or someone else) has talked about microwaving vials.


----------



## Tony121

angus_grant said:


> I've only done a few batches so not really commenting from experience. Haven't actually roused any yeasts from frozen sleep yet, so grain of salt....
> Microwaved for 10 mins. Not melted any vials yet.
> Pretty sure yob (or someone else) has talked about microwaving vials.


Just to confirm, the solution is in the vials when you microwave or do you do that separately?


----------



## angus_grant

Yep glycerine/water in vial. 
Cool vial down before adding yeast to avoid cooking your yeasties. 
Just replace step 4 of Mardoo instructions with microwaving


----------



## Tony121

Thanks Angus.

Thought I'd try out with rinsed yeast from my last batch. Still need to refine technique but got there in the end; 

Lid too loose = overflow in microwave
Lid too tight = bang (yes I know obvious, but microwave needed a clean anyway so thought I'd give it a bash and see what happened)

Anyway, will see how they end up in a few weeks/months time.


----------



## Schooner_downunder

Just starting to get into yeast freezing but just wanted to check if there is any difference between vegetable and petroleum based glycerine.


----------



## Mardoo

You want vegetable.


----------



## Midnight Brew

Wyeast 3068 destined for a dunkelweizen.

Date frozen : [SIZE=11pt]*24.01.2014*, 10% Glycerine[/SIZE]

*3 years to the day.*

100ml 1020 starter for 48-72 hours ---> 1L 1040 starter for 24 hours


----------



## SBOB

I revived my first frozen vials which was a 7month old wyeasy1056

0.5L 1.020 for 24 hours then 2L 1.040 and was decent activity after another 24 hours

Are those steps the 'recommended' or should I adjust volumes/step sizes?


----------



## Moad

thats what I run with mate, I then decant most of the 2L starter and spin up another 2L @ 1.040 for double batches


----------



## Mardoo

That's exactly what I've been doing. It's only taking two steps, which has been a surprise. Not sure whether it's rushing it. Wasn't intending to do two steps. My small flask was dirty so I just did a 500ml first step followed by 2L. I've been getting just what I need for a single pitch. Only done it twice. 

Hard to tell whether viability has been affected, as the wort I've fermented the pitches on had a very unpredictable aspect in its production. Any opinions or information on the effect of two steps vs three?


----------



## Lord Raja Goomba I

And what ratio became the norm?

Water to Glycerine ratio (50/50 water to glycerine?). Or expressed as 37.5/37.5/25 ratio of water/glycerine/yeast slurry? Just for my accountant pedantic brain.

I've just grabbed a 1L for $7.95 plus delivery evilbay vegetable glycerine amount and will do this on the weekend.

(BTW I absent mindedly froze some WLP001 without doing this, so might resurrect it and see if it fires and then can be split. Any thoughts?)


----------



## kaiserben

Lord Raja Goomba I said:


> I've just grabbed a 1L for $7.95 plus delivery evilbay vegetable glycerine amount and will do this on the weekend.


Could you provide the link for this?


----------



## Lord Raja Goomba I

kaiserben said:


> Could you provide the link for this?


http://www.ebay.com.au/itm/191613432703


----------



## malt junkie

LRG, I do 50/50 glycerine/ water to half vial and top the rest with yeast in a 50ml vial. works for me!


----------



## Lord Raja Goomba I

malt junkie said:


> LRG, I do 50/50 glycerine/ water to half vial and top the rest with yeast in a 50ml vial. works for me!


Sweet, thanks heaps.

Love to hear more confirmation stories.


----------



## Yob

Same ratio for me, better, ot least less harsh environment and the yeast seem to be able to expel the glycerine faster than a higher ratio


----------



## Lord Raja Goomba I

Good, so that ends up being a 25/25/50 ratio of water, glycerine and yeast?


----------



## Midnight Brew

Lord Raja Goomba I said:


> And what ratio became the norm?
> 
> Water to Glycerine ratio (50/50 water to glycerine?). Or expressed as 37.5/37.5/25 ratio of water/glycerine/yeast slurry?
> 
> (BTW I absent mindedly froze some WLP001 without doing this, so might resurrect it and see if it fires and then can be split. Any thoughts?)


Originally I used a 10% ratio of glycerine. It works great when frozen correctly. At 10% you need to crash the yeast to below freezing temperatures as fast as possible. If you just pop it in the freezer and let it freeze slowly the cells are more likely to shear. I had a conversation with Grainer about it a few years back when I first started freezing. There was another reason but I cant quiet recall. Grainer has a background in microbiology.

At 25% the cells dont actually freeze but more or less stay suspended in time. Less chance of shearing the cells. And just about everyone that has used it at this percentage has reported back with good results.

As for your WLP001. I doubt it will resurrect but please report back your results.


Get ready to dive into the hobby that is yeast farming. May your yeast be fruitful and multiply.


----------



## Lord Raja Goomba I

Midnight Brew said:


> Originally I used a 10% ratio of glycerine. It works great when frozen correctly. At 10% you need to crash the yeast to below freezing temperatures as fast as possible. If you just pop it in the freezer and let it freeze slowly the cells are more likely to shear. I had a conversation with Grainer about it a few years back when I first started freezing. There was another reason but I cant quiet recall. Grainer has a background in microbiology.
> 
> At 25% the cells dont actually freeze but more or less stay suspended in time. Less chance of shearing the cells. And just about everyone that has used it at this percentage has reported back with good results.
> 
> As for your WLP001. I doubt it will resurrect but please report back your results.
> 
> 
> Get ready to dive into the hobby that is yeast farming. May your yeast be fruitful and multiply.


Thanks bud. Looking forward to this - fun wise and cost wise (and having a good bank of something other than dried yeast).

I've done a bit of research and apparently, there may be 10% or so of viable yeast in a frozen smack pack. In the US, they have this stuff called snow and sometimes they get stuff delivered and it's frozen because of it. So a starter and they can get it moving.

I wish we got snow. I hate the heat.


----------



## Mardoo

Again, another for the 25/25/50 ratio. Works the best of the the three I've tried, being 10% glycerine, 25%, and 50%. Don't bother with 50%...


----------



## Yob

Anecdotally, the %50 is harder for the yeast to expel, seems like it takes longer for the yeast to fire up in the starter, but it stays liquid and doesn't freeze solid, I feel the trade off to a %25, while it does go solid is a better mix with no noticeable drift after over 2 years in the deep freezer. Last count I had over 180 vials and over 30 strains...


----------



## Moad

another 25/25/50% here. I don't treat my yeasties too well, They are just in a container in a frost free freezer with one of those freezer gel packs thrown in. I have had no issues whatsoever with spinning up yeast (granted only up to 8 or 9 months so far) and achieving great fermentation indiscernible from starter made straight from smack pack.

This has possibly been the greatest thread I have been involved with in my AHB career! Thanks to Yob especially for continued contribution

p.s when are the vaults going into full production?


----------



## Yob

I'm deciding over the vault walls, will post in the other thread as to the direction.


----------



## MartinOC

I'm quivering in anticipppppppation!


----------



## Yob

it's terribly addictive...

its also a fascinating sub hobby to brewing.. and even a hobby in its own right...


----------



## malt junkie

But if you weren't brewing what good would yeast freezing be? Ok you could start baking but how many strains could you actually bake with?
I feel a yeastperiment may be on the cards. Hefe-danish anyone?


----------



## peteru

Beer yeasts are too slow for most baking needs. I tried making pizza with Cooper's kit yeast. After making one biscuit with pizza toppings, I just got out the dried bakers yeast, teaspoon of sugar and some water and fixed up the dough. Night and day. The rest of the pizzas turned out fine.


----------



## Mardoo

Actually many brewpubs in the States use hefeweizen yeasts for their pizza dough. Maybe it's the massive krausen ones that would work better.


----------



## Schooner_downunder

Can someone let me know if I have stuffed up something with some yeast I recently tried to freeze. Should Glycol be on top or bottom?

My process was I started with a White labs pack of WLP001 (California Ale) and a 500 ml starter for 12 hours, then stepped to 1.5 liters and ran for 36 hours to high Krazen. Put into the fridge overnight to settle yeast decanted and put a 50/50 mix into 50 ml tubes.

My fridge goes down to - 23 C. Here is a pic of the yeast. I may be paranoid but is the vegetable glycol suppose to be sitting on the bottom and the yeast on the top?


----------



## kcurnow

Nope should be fully mixed


----------



## Yob

Also, where'd you get that tray?


----------



## Schooner_downunder

Brewnut said:


> Nope should be fully mixed


Damn!! I may tried to bring back to refrigerator temperature and shake to mix. Worse case it all came from the same pack so not out of pocket that much... fingers crossed I can recover something


----------



## Schooner_downunder

Yob said:
 

> Also, where'd you get that tray?


Ebay - Blue Plastic 21 Holes Box Rack Holder for 50ML Centrifuge Tubes WS

http://www.ebay.com.au/itm/Blue-Plastic-21-Holes-Box-Rack-Holder-for-50ML-Centrifuge-Tubes-WS-/222137295119?hash=item33b86a210f:g:3GQAAOSwvg9XTpdQ


----------



## Yob

Schooner_downunder said:


> Ebay - Blue Plastic 21 Holes Box Rack Holder for 50ML Centrifuge Tubes WS
> 
> http://www.ebay.com.au/itm/Blue-Plastic-21-Holes-Box-Rack-Holder-for-50ML-Centrifuge-Tubes-WS-/222137295119?hash=item33b86a210f:g:3GQAAOSwvg9XTpdQ


that sir, is ******* excellent work

going to order 15


----------



## Yob

oh shit.. I just felt the bulk buy tingle...

#shakesfistatschooner


----------



## Lord Raja Goomba I

Final question,

the 50% yeast slurry. Is that yeast with a little bit of worth that has been mixed into it? Or just the pure yeast.

Reason I asked, is I"m looking at the Splitting a smack pack thread, and I fired up a smack pack that was old and I thought stuffed, and now it's working and is sitting on 1L of 1.040 starter.

Do I ferment out and nick the pure yeast?


----------



## Mardoo

I always have enough wort or sterile water to make the yeast liquid enough to work with easily. If splitting straight from a fresh smack pack, I split it into 5 50ml vials (including the glycerine solution).


----------



## Lord Raja Goomba I

Gents, any place you get test tubes in Australia? 

I reckon my 10 tubes will be only enough for one batch of yeast I've been culturing for a week and I don't want to wait until the baby reaches puberty until I get the Guangzhou specials.


----------



## Schooner_downunder

Lord Raja Goomba I said:


> Gents, any place you get test tubes in Australia?
> 
> I reckon my 10 tubes will be only enough for one batch of yeast I've been culturing for a week and I don't want to wait until the baby reaches puberty until I get the Guangzhou specials.


I bought these 50 ml autoclavable tubes (LCAO) from Proscitech (Mentioned earlier in the thread) and they work like a charm. I got 50 for $ 21 + Shipping, happy to ship a few your way if you want to test them out.

https://proscitech.com/?navaction=show_page&chapter=l&page=9#lca01


----------



## Lord Raja Goomba I

Schooner_downunder said:


> I bought these 50 ml autoclavable tubes (LCAO) from Proscitech (Mentioned earlier in the thread) and they work like a charm. I got 50 for $ 21 + Shipping, happy to ship a few your way if you want to test them out.
> 
> https://proscitech.com/?navaction=show_page&chapter=l&page=9#lca01


That would be awesome! Thanks so much! 

I just found out that the Guangzhou specials don't like being heated to boiling water temps for an hour. The caps held up, though.


----------



## SBOB

Lord Raja Goomba I said:


> That would be awesome! Thanks so much!
> 
> I just found out that the Guangzhou specials don't like being heated to boiling water temps for an hour. The caps held up, though.


really?
The ones I got from ebay go fine in the pressure cooker

Different seller, but same as these
http://www.ebay.com.au/itm/10X50ml-Plastic-Centrifuge-Test-Tubes-Vial-Container-Self-Standing-Screw-Caps-/272438554310


----------



## Lord Raja Goomba I

SBOB said:


> really?
> The ones I got from ebay go fine in the pressure cooker
> Different seller, but same as these
> http://www.ebay.com.au/itm/10X50ml-Plastic-Centrifuge-Test-Tubes-Vial-Container-Self-Standing-Screw-Caps-/272438554310


Looks like on further research I didn't check if they were autoclavable.


----------



## SBOB

Lord Raja Goomba I said:


> Looks like on further research I didn't check if they were autoclavable.


rookie mistake


----------



## Camo6

Even the autoclavable ones don't like being pressure cooked with the lids screwed down tight. Another rookie mistake.


----------



## Mardoo

Yeah, definitely leave the lids LOOSE when you pressure cook vials, otherwise the sides cave in once they get hot.


----------



## Lord Raja Goomba I

Camo6 said:


> Even the autoclavable ones don't like being pressure cooked with the lids screwed down tight. Another rookie mistake.





Mardoo said:


> Yeah, definitely leave the lids LOOSE when you pressure cook vials, otherwise the sides cave in once they get hot.


Thanks for the tips, gents. I took the lid off mine, and whether that hurt the cause, I can't think it would have made that much difference to the body shape, but who knows?

Looking at prositech, I'm thinking bulk buy for SEQ? Free delivery for over $250 orders and I reckon a little negotiating might get a cheaper per unit price.


----------



## Lord Raja Goomba I

Another stoopid kitten in airlock, RDWHAHB type of question.

My frozen yeasts appear to have slightly separated. By which there appears to be a denser brown liquid at the bottom and the balance being 'yeast brown', but not so dense looking. 

I shook them up before chucking them in the deep freeze (which is not frost free) to assimilate the glycerine/water and yeast. Used the 25/25/50 ratio.


----------



## SBOB

Lord Raja Goomba I said:


> Another stoopid kitten in airlock, RDWHAHB type of question.
> 
> My frozen yeasts appear to have slightly separated. By which there appears to be a denser brown liquid at the bottom and the balance being 'yeast brown', but not so dense looking.
> 
> I shook them up before chucking them in the deep freeze (which is not frost free) to assimilate the glycerine/water and yeast. Used the 25/25/50 ratio.


based on my experience, perfectly normal

All my frozen samples still settle before freezing so yeast/trub at bottom and clearer liquid at top


----------



## Mardoo

Same here.


----------



## sp0rk

If you're going to get them from Proscitech, get the LCA03-50
My LCA01 free standing tubes sometimes leak if knocked over


----------



## Lord Raja Goomba I

sp0rk said:


> If you're going to get them from Proscitech, get the LCA03-50
> My LCA01 free standing tubes sometimes leak if knocked over


Bugger. I grabbed the LCA01, they looked the goods.

I haven't yet seen any leaks and I whacked one in the fridge to do a mild that was relegated because I needed some more AIPA, so I'll see how that goes. I have an empty six pack from a Carslberg special that holds them up when in the freezer.

Good to know though.


----------



## SBOB

sp0rk said:


> If you're going to get them from Proscitech, get the LCA03-50
> My LCA01 free standing tubes sometimes leak if knocked over


$4... problem solved 
http://www.ebay.com.au/itm/222262953615


----------



## Lord Raja Goomba I

SBOB said:


> $4... problem solved
> http://www.ebay.com.au/itm/222262953615


Good spot.

I think SWMBO is cottoning on to the fact that I make smaller purchases regularly to slip under the Minister of War and Finance Radar.


----------



## Yob

I've got one on the way too, they look the goods


----------



## Gloveski

Silly newbie question put could you use sanitised zip lock bags . I presume there would be some form of sterisation as they are mainly used for food . Only real issue I could see would be pouring them into a starter


----------



## kcurnow

I have also ordered a couple, but I think they must be coming by carrier pigeon that's sitting on a container ship as the expected arrival date is showing as may!! Be wary of free shipping as while it might be free it can take a while.


----------



## Mardoo

Mine said April but came a few weeks ago. Fret not.


----------



## Yob

Gloveski said:


> Silly newbie question put could you use sanitised zip lock bags . I presume there would be some form of sterisation as they are mainly used for food . Only real issue I could see would be pouring them into a starter


No


----------



## SBOB

Yob said:


> I've got one on the way too, they look the goods


mine already arrived
Fits the vials without issue (pretty firmly)

For $4 (or in your case, about $400 for all your vials) its a good value vial rack


----------



## Camo6

Looks like Yob has already placed his order as they were out of stock when I looked.


----------



## Yob

Asked for pricing on 200 but think I scared him a little..


----------



## mofox1

Another success story for the thread... WLP004 I froze from the original white labs vial in Jun 2015.

5ml yeast solids in the vial and after 3 steps / 5 days I had a 2.5L starter starting to drop clear at ambient temps. Not a bad result from 1/6 of a yeast purchase and 2$ sugaz.


----------



## Yob

best value thread on the interwebs...


----------



## kaiserben

Stupid question time: 

How do you get your liquids into these little vials?


----------



## mofox1

Pouring carefully?

Seriously though, I'm either filling vials from fresh packs of yeast or from narrow mouthed flasks. Even if I rinse a yeast cake in larger jars, it ultimately ends up in a flask on (after?) the final rinse. Although I hardly bother with yeast rinsing any more... it's just not worth the effort when I can spin up another dozen vials worth on the stir plate.


----------



## Yob

What he said.. So simple


----------



## SBOB

yep
pour gently from Erlenmeyer flask.. even with a wide flask its easy after the first couple of tries

decant as much spent wort as possible leaving as thick a yeast cake as you can, then pour into vials which already have been prepared (water/glycerine combo + pressure cooked)


----------



## kaiserben

I wouldn't have any trouble pouring from fresh packs (because you can bend the packaging so it forms a spout), but I was a bit worried about pouring from my 3L Erlenmeyer flask. 

EDIT: And my plan was to make starters from fresh packs (which often aren't all that fresh after sitting in a shop for months), chill and decant wort, then get yeast into vials. Anyway, I'll see how it goes.


----------



## Lord Raja Goomba I

I pour from my erlenmeyer into a glass jar to settle off the remaining wort and give it a slight rinse - nothing exhaustive. From there I use the syringe trick that BribieG outlines at the start.

That also clears up the erlenmeyer for the next spin.


----------



## malt junkie

I use a beaker then a huge syringe, well 2 syringes one for yeast one for glycerin mix.


----------



## kaiserben

Ah cool, just re-read this thread and saw the syringes in the first post. 

What syringes are you guys using? (are they autoclavable? If not, what are you doing to sanitise them?)


----------



## malt junkie

Chemist... well not one in a small town but most big chemists should carry some decent size ones, I just throw them is the pressure cooker when I do a couple batches of vials. I have two 50ml need to track down some spares though.


----------



## Yob

Started with that, don't use them bow as it's just more risk


----------



## kaiserben

I might try pouring from my Erlenmeyer into a pressure cooked pyrex jug (which has a spouty-type thing to make pouring somewhat easier) and then from there into the vials. 

Right-e-o, I've just bought a bunch of stuff: 

I bought 3 x 21 hole rack vial holders: link for AU$11.26 delivered. 
2 x 1kg Vegetable Glycerine: link for AU$30.85 inc delivery. 
4 x 10 centrifuge vials: link for AU$22 delivered (was gonna get this from Proscitech, was a bit unsure if I wanted to pay $15 on shipping for $25 worth of stuff (ie $40 total for 50 vials/LCA03), eventually decided I'd buy anyway, but then discovered they still had to add GST on top AND were charging GST on the shipping. Only added $4 more, but still ...)


----------



## Tony121

Random question, does the glycerine break down or go off if you keep it for an extended period of time?

Reason I ask is that I bought a 5kg bottle for about $25 and can't see me using it all for some time.


----------



## kaiserben

By the time a yeast pack gets to me it's usually a few months old (sometimes older, rarely younger).

According to YeastCalc 3 months after the manufacturing date means the original 100bn viable cells are more like 46bn. 

So as an example, assuming I have a "fresh" pack of yeast with 46bn viable cells, what is everyone's opinion on whether it's better to: 

1. Split the fresh pack of yeast into vials as is. 
or
2. Do a starter to build the cells up a bit and revitalise the cells. 

I realise there is a bit of a trade-off, where it would no longer be a pure/1st-gen culture once you've performed a starter, but at least the yeast you're freezing will be healthy and in good numbers.

What are people's thoughts on this?


----------



## peteru

You could do both. Archive 1/2 the pack and make a starter from the other half. You can compare the results down the track and decide which method you prefer.


----------



## Mardoo

I'll grow the pack up in that situation, just to ensure the yeast is well-fed and healthy and ready for the big sleep.

Another solution is to get onto Brewman's Wyeast pre-ordering buys. The yeast comes fresh as! You may still be able to squeak in on the current one, but hurry. 

Also, you can contact the other forum supporters and find out when their next order is and see whether you can pre-order.


----------



## kaiserben

Yeah - I've recently jumped in on an AHB Yeast Bay bulk buy, and also just pre-ordered a couple of strains from the White Labs Vault.


----------



## Mardoo

How'd you do the Yeast vault pre-order?


----------



## kaiserben

Via my LHBS. (Dunno if they're a forum sponsor though) 

Check this forum thread: http://aussiehomebrewer.com/topic/94457-white-labs-wlp059-melbourne-ale-yeast-to-be-released/


----------



## kaiserben

And this info from White Labs: http://www.whitelabs.com/news/the-vault-hits-production-2017


----------



## Meddo

Looking to start some yeast freezing shortly, I'm gradually working my way through this thread but haven't yet found much mention of splitting White Labs Purepitch packs as opposed to Wyeast smack packs. The White Labs packs only have ~40 mL of yeast/slurry compared to the 125 mL in a Wyeast smack pack.

Given that it seems the consensus is to go 25% water / 25% glycerin / 50% yeast slurry, I've got 50 ml vials, and would like to get 5 vials worth from a Purepitch pack, would the best practice option be:

1 - 4 mL water / 4 mL glycerin / 8 mL yeast ( 16 mL total in 50 mL vial, 25/25/50 )
2 - 21 mL water / 21 mL glycerin / 8 mL yeast ( 50 mL total in 50 mL vial, 42/42/16 )

or just grow up a starter and go:

3 - 12.5 mL water / 12.5 mL glycerin / 25 mL yeast slurry from starter ( 50 mL total in 50 mL vial, 25/25/50 )

It seems from what I have read that the 50/50 water to glycerin mix is optimal, and the mix volume should match or exceed that of the yeast? If that's the case then the first two options really only differ in the amount of headspace in the vial and how quickly I burn through my glycerin stash, so is there any downside to having 30+ mL of headspace in a vial as per option 1? (apart from inefficient use of freezer space)

If 8 mL of yeast won't be enough to easily culture up upon thawing (for a yeast-wrangling beginner) then option 3 gives me a full 25 mL yeast in each vial but presumably with an increased likelihood of infection.

Thoughts?

Thanks (and apologies if this has already been answered)


----------



## Lord Raja Goomba I

Meddo said:


> Looking to start some yeast freezing shortly, I'm gradually working my way through this thread but haven't yet found much mention of splitting White Labs Purepitch packs as opposed to Wyeast smack packs. The White Labs packs only have ~40 mL of yeast/slurry compared to the 125 mL in a Wyeast smack pack.
> 
> Given that it seems the consensus is to go 25% water / 25% glycerin / 50% yeast slurry, I've got 50 ml vials, and would like to get 5 vials worth from a Purepitch pack, would the best practice option be:
> 
> 1 - 4 mL water / 4 mL glycerin / 8 mL yeast ( 16 mL total in 50 mL vial, 25/25/50 )
> 2 - 21 mL water / 21 mL glycerin / 8 mL yeast ( 50 mL total in 50 mL vial, 42/42/16 )
> 
> or just grow up a starter and go:
> 
> 3 - 12.5 mL water / 12.5 mL glycerin / 25 mL yeast slurry from starter ( 50 mL total in 50 mL vial, 25/25/50 )
> 
> It seems from what I have read that the 50/50 water to glycerin mix is optimal, and the mix volume should match or exceed that of the yeast? If that's the case then the first two options really only differ in the amount of headspace in the vial and how quickly I burn through my glycerin stash, so is there any downside to having 30+ mL of headspace in a vial as per option 1? (apart from inefficient use of freezer space)
> 
> If 8 mL of yeast won't be enough to easily culture up upon thawing (for a yeast-wrangling beginner) then option 3 gives me a full 25 mL yeast in each vial but presumably with an increased likelihood of infection.
> 
> Thoughts?
> 
> Thanks (and apologies if this has already been answered)


Hey Meddo, I've just done this twice, let me have a chance to formulate "what I did" and I"ll post back.


----------



## Lord Raja Goomba I

[SIZE=10pt]I had two WLP 'pitchable yeast' packs.[/SIZE]

[SIZE=10pt]One was out of date (a freebie from a nice retailer) and one was in date.[/SIZE]

[SIZE=10pt]For the In date one, I built a starter, starting with 500ml at 1.035-ish, on the stir plate for 24 hours and then pitched another 1L at the same SG straight in. This took about 24 hours more, and after cooling, I was ready to freeze in about 3 days (though I took an extra day to stuff around and get to it).[/SIZE]

[SIZE=10pt]I ended up with 6 vials (50ml vials, so 25ml of yeast per vial) and froze 5 using the 25/25/50 ratio of water/glycerine/yeast.[/SIZE]

[SIZE=10pt]I pitched the 6th just this morning, so I'll let you know how that goes.[/SIZE]

[SIZE=10pt]With the out of date one, this took (understandably) longer.[/SIZE]

[SIZE=10pt]I built a 1L starter at 1.020. Waited for it to ferment out in 2 days, cooled, decanted the wort, 2L at 1.020, repeat the ferment and cool and decant. Then 1.0L at 1.040 of wort for another 2 days, decanted, and another 2L at 1.040.[/SIZE]

[SIZE=10pt]From this, I managed 5 vials and froze these using the above 25/25/50 ratio.[/SIZE]


----------



## Lord Raja Goomba I

And it's off. 2 inches of Krausen on a low grav beer (and I kinda abused the non-frozen yeast, by leaving it in the fridge sans-glycerine and no starter for a couple of weeks).

I'm impressed.


----------



## kaiserben

Yob said:


> re-activating: Are you guys cold crashing between steps or just adding the 200ml to the 500ml of fresh wort etc? (I'll make up the next step to include the volume.. so, if I have a 500ml starter, Ill make up a 200g/1.5lt starter and add the 500ml to make the full 2000ml if that makes sense?)
Click to expand...

This little bit of info from Yob has really streamlined my starter-steps process. 

It might already have been obvious to a lot of people, but it wan't to me. Previously I'd been crash chilling between every step when doing multi-step starters, ditching spent wort and the whole process was time-consuming and fiddly. But this method is way easier, saves heaps of time and gives me more options.


----------



## mofox1

kaiserben said:


> This little bit of info from Yob has really streamlined my starter-steps process.


Yup. He's a keeper, that's for sure (sorry, "fo' sho").


----------



## Yob

Fo shoz?


----------



## kaiserben

I assume it's okay to re-use your vials? And, if so, how do you label them?

And also, I've now got some racks for my vials. I'm looking for a suitable box to house them. How about a little esky with some ice bricks alongside the racks of vials?


----------



## Yob

I write on the lid so I can identify easily 

Fine Re use, boil sanitiser, done


----------



## SBOB

i use small stickers on top to right on.. peel off when washing for vial re-use


----------



## Yob

and if you have a frost free, certainly use ice bricks.. I dont use anything in the deep freezer though


----------



## SBOB

SBOB said:


> i use small stickers on top to right on..


Wow..that should obviously be 'write on'... Though perhaps the stickers are for encouragement like when you reward children, and I subconsciously meant that I say 'right on' to cheer on my yeast..


----------



## Moad

I number mine and track in google sheets with other inventory. I also made a recipe builder that deducts from inventory. Yes I like spreadsheets


----------



## Mardoo

Hey Melbourites,

Started a yeast vial mini-BB thread if you want/need more vials:

http://aussiehomebrewer.com/topic/94604-e-melbourne-yeast-vials-mini-buy/


----------



## lfc_ozzie

Probably a dumb question, but is there a difference between getting a stove top pressure cooker and an electrical pressure cooker? or does it not matter which one you use to boil the 50/50 glycerin/water mix in? 

Can't do the microwave method as i don't have one, currently i just boil my vials in a pot for 15 minutes to sterilize and store yeast in these with cooled boiled water in the fridge for future use, but want to get into freezing as they will last longer and seems more fun


----------



## Mardoo

Doesn't matter stovetop or electric.


----------



## lfc_ozzie

Mardoo said:


> Doesn't matter stovetop or electric.


Thanks mate


----------



## Gloveski

well I have just froze my first lot of yeast M76 Bavarian Lager .................thinking of having a go at drying some yeast also down the track , I think I'm addicted to all things homebrew lol


----------



## kaiserben

2 weeks ago I froze my first yeast.

Already I have 5 different strains (and a total of 20 vials) in the freezer. 

Soon I'll be out of vials and freezer space ...


----------



## MartinOC

There's always this if you're up for postage: http://aussiehomebrewer.com/topic/94604-e-melbourne-yeast-vials-mini-buy/#entry1445414


----------



## Yob

kaiserben said:


> 2 weeks ago I froze my first yeast.
> 
> Already I have 5 different strains (and a total of 20 vials) in the freezer.
> 
> Soon I'll be out of vials and freezer space ...


good work... do you need an 1100lt deep freezer? h34r: :lol: 

(only kidding, I need it for the next brewman yeast buy up)


----------



## labels

Just a thought, this might already have been discussed in this thread but too long to back through it all - will food grade propylene glycol work as well as glycerine??

The reason I ask is the way I understand things, the yeast survive freezing temperatures because the glycerine doesn't freeze and prevents ice crystals from bursting the yeast cells.

I don't suppose it matter which one you use, both are water soluble and both can get well below freezing, I think it's -50C for PG


----------



## Tony121

My understanding is that the yeast absorb the glycerine which stops them from bursting somehow (someone with more knowledge than I will know). Mine completely freeze though fire up without any issue.

I could be well truly wrong on this one.


----------



## Mardoo

There are two schools of thought I've run across. One is that the glycerine keeps the yeast from freezing. The other is that the glycerine keeps the yeast from dehydrating. However, that doesn't answer the question of whether glycerine or propylene glycol. Give it a try. do yeast from the same batch, half with glycerine, half with pg. Be interesting to see how it goes.


----------



## Yob

my thoughts lead toward the glycerine expelling the water, when you pour some in the vial it displaces the water, if the yeast membrane still had water it would still freeze right?

no science behind this, just the way I think it would be, the yeast is still at -20 in the deep freezer so any water will surely be ice crystal?


----------



## Gloveski

keeping it simple going to freeze some US05 and use this first so I can easily use a dried packet if it fails .
Haven't done a starter yet still waiting on a yeast forge , may try one without a stirplate while I wait .
Once I have success will be hitting Brewman up for some different yeast strains


----------



## Yob

That's the spirit


----------



## Yob

In no way do I have a problem.... even if I am doing a major upgrade to my yeast bank


----------



## Lord Raja Goomba I

Thanks for bringing this to recent threads, Yob.

I'm going to take possession of a wlp funky cider yeast. I want it to last.

I assume I use apple juice as a starter, to multiply it to get the mandatory 5 vials from one tube.


----------



## Yob

Yep, use the medium that it's to actually ferment, added yeast nutrient though


----------



## Curly79

Yob said:


> In no way do I have a problem.... even if I am doing a major upgrade to my yeast bank
> 
> 
> 
> 
> yeast vilas 1.JPG
> 
> 
> 
> 
> yeast vilas 2.JPG


Maybe you should have hung onto that chest freezer mate


----------



## Lyrebird_Cycles

labels said:


> Just a thought, this might already have been discussed in this thread but too long to back through it all - will food grade propylene glycol work as well as glycerine??
> 
> The reason I ask is the way I understand things, the yeast survive freezing temperatures because the glycerine doesn't freeze and prevents ice crystals from bursting the yeast cells.
> 
> I don't suppose it matter which one you use, both are water soluble and both can get well below freezing, I think it's -50C for PG





Mardoo said:


> There are two schools of thought I've run across. One is that the glycerine keeps the yeast from freezing. The other is that the glycerine keeps the yeast from dehydrating. However, that doesn't answer the question of whether glycerine or propylene glycol. Give it a try. do yeast from the same batch, half with glycerine, half with pg. Be interesting to see how it goes.



Glycerol will work better than polypropylene glycol. The way glycerol works is not intuitive: it's apparently due to the very complete mixing of glycerol and water inhibiting the ability of either of them to link up, thus inhibiting crystal formation. Reference here.

Simple freezing point depression (eg propylene glycol) is a different mechanism: it's a colligative property, meaning that it depends on the quantity of solute present not on its identity. In the case of freezing point depression it works by diluting the solvent (water) thus reducing its equilibrium vapour pressure via Raoult's Law. This in turn reduces the temperature at which the vapour pressure of the liquid equals the vapour pressure of the crystal form (ice) which is the definition of freezing point.


----------



## lfc_ozzie

Hi guys,

Finally got around to freezing some yeast, only did two vials as a test for now as i used the rest of the starter in a beer. Anyway i have attached photos, do these look ok for frozen yeast? Any feedback is appreciated


----------



## Lord Raja Goomba I

Looks about what mine does.

It separates still, but I've had no issue with thawing and kickstarting them.


----------



## Mardoo

Totally normal.


----------



## AJS2154

Yob said:


> In no way do I have a problem.... even if I am doing a major upgrade to my yeast bank
> 
> View attachment 97147
> 
> 
> View attachment 97148


None of us have a problem, Yob. Until we admit we have a problem all is well.

I have a question though. With a bank of that magnitude, I imagine the stock turns are relatively low. If that is the case some of your yeast must sit in the freezer for some time. Have you determined an optimum use by shelf (freezer) life? I have some that are about 10-12 months old at times and they seem to fire up OK.

See you, Anthony


----------



## MartinOC

Lyrebird_Cycles said:


> Glycerol will work better than polypropylene glycol. The way glycerol works is not intuitive: it's apparently due to the very complete mixing of glycerol and water inhibiting the ability of either of them to link up, thus inhibiting crystal formation. Reference here.
> 
> Simple freezing point depression (eg propylene glycol) is a different mechanism: it's a colligative property, meaning that it depends on the quantity of solute present not on its identity. In the case of freezing point depression it works by diluting the solvent (water) thus reducing its equilibrium vapour pressure via Raoult's Law. This in turn reduces the temperature at which the vapour pressure of the liquid equals the vapour pressure of the crystal form (ice) which is the definition of freezing point.



I have no doubt that you know what you're talking about, but you completely lost me here with the science.


----------



## Yob

AJS2154 said:


> None of us have a problem, Yob. Until we admit we have a problem all is well.
> 
> I have a question though. With a bank of that magnitude, I imagine the stock turns are relatively low. If that is the case some of your yeast must sit in the freezer for some time. Have you determined an optimum use by shelf (freezer) life? I have some that are about 10-12 months old at times and they seem to fire up OK.
> 
> See you, Anthony



Ive just used some of my oldest at over 3 years.. shit, you'd have to check the start of the thread where I started but I revived one of those and used it for 6 successive pitches over 3 styles...

crazy good shit


----------



## Yob

Posted 5/1/14

There you go.. 3 1\2 years roughly


----------



## AJS2154

Mate that is amazing. They have to be one of the best organisms on this earth.......tough, work hard, and create a beautiful product. Hard not to admire them. 

See you, Anthony.


----------



## Gloveski

Well first crack at a starter from frozen lager yeast initially started with 500ml , had to fridge it for a week then made a new starter last night and already a nice healthy krauzen. Just need a stirplate now , hopefully digital homebrew wont be to far off with the new model


----------



## Nullnvoid

How long before adding the yeast do people sterilise the tubes with the water glycerine mixture? Will a couple days/week be alright?


----------



## kaiserben

Nullnvoid said:


> How long before adding the yeast do people sterilise the tubes with the water glycerine mixture? Will a couple days/week be alright?



I think I remember reading somewhere that up to a couple of weeks is fine. 

I tend to do it a day or a few days ahead (then store in fridge till the yeast is ready). 

If I end up with spares I keep them in the fridge and then re-sterilise a few days before the next time.


----------



## Nullnvoid

So I have successfully (I hope) just put my first batch in the freezer! 

I have another vial in the fridge to freeze down. 

Question is, do I freeze this first batch for a couple days and then try and restart one to see if it works or do I immediately jump onto the next one to be frozen?


----------



## Mardoo

You can have a quick re-start if you want, but if you followed the processes we all do (presumably  ) it will work.


----------



## Nullnvoid

Yeah followed the "official" unofficial accepted procedure. 

Just not sure I drained enough wort off before I filled the vials. But I guess as long as there is some yeast in there, it should be right.


----------



## Mardoo

I drain my wort off until there's enough wort left to make the wort/yeast mixture about as thick as somewhere between undiluted cordial and motor oil. Whatever the case, there WILL be yeast in there. The only difference might be how many steps you need to get a big enough pitch. 

Wouldn't hurt at all to start one up. It's something you get a feel for after a few batches.


----------



## Nullnvoid

Thanks Mardoo! Very helpful. I think I'll restart one to be sure in my own mind. I have a cube ready to go anyway so it would be timely anyway.


----------



## Nullnvoid

The next question I have for the brains trust is with the freezing. 

I have limited room in the freezer and have managed to score a small polystyrene container. I tried freezing some water in it with vials with the thought I could pull the vials out and then just use the holes with ones filled with yeast but this wasn't as successful as I'd liked. Couldn't easily get the temporary vials out and then put the new ones in and now checking they are hard too. 

Is the polystyrene container enough to maintain again the defrost cycle or what does everyone else do?


----------



## Mardoo

I just chuck a gel ice pack or two on top of the vials. I've tried a number of mediums like you did with ice, and, so far, all have been more trouble than they're worth. A hell ice pack on top of the vials inside a closed container has worked great for me. If the container is insulated, even better.


----------



## Nullnvoid

Sweet. Sounds like I'm making it harder than it needs to be and over thinking it .


----------



## koshari

such an excellent guide, i pdf bombed this before the photobucket images get toasted.


----------



## Mardoo

koshari said:


> such an excellent guide, i pdf bombed this before the photobucket images get toasted.


Care to post it for posterity?


----------



## koshari

Mardoo said:


> Care to post it for posterity?


yeah i will , just need to sort out some storage options, seems the forum posting world has been pretty much rocked a bit by the photobucket debacle.


----------



## kaiserben

Having now banked 10 strains, tonight I'm finally going to revive some (WLP029).

If I'm looking at building to 270bn cells, what's the best way of figuring out how to get there and how many cells I've got at any given point? Is the only realistic way by doing a starter and then looking at the amount of slurry in my flask to try to guesstimate how many cells I end up with?


----------



## Mardoo

Use the CaptainBrew pitch rate calculator. It allows you to do up to three steps-up. There are various options for aeration. If using a stir-plate I recommend the Kai Troester setting. People differ on this. That gives me an amount that always gives me clean ferments that don't stall. You can also calculate DME or LME amount to use, if you use it. 

https://www.captainbrew.com/yeast-pitch-rate-and-starter-calculator

The only way to know count and health for sure is to use a high-powered microscope and a cytometer to count by hand and then estimate up. For homebrewing purposes you don't need to geek that hard, unless YOU need to geek that hard. I may do so in the future, but I just use broad volumetric estimates and experience to work out yeast count and health. 

For the first step up, use 1.020 wort, just to give the yeast a gentle wake up. For ales I can usually get to pitching volume in two steps. You can adjust all those settings in the Captain Brew calculator, as well as initial pitch, which I estimate as % of original yeast pack into the vial - 20% to allow for death from freezing, I use the number of weeks from the production date to when I froze to estimate viability. 

It's something you'll grow used to as you do it over time. Hope that all makes sense. Open up the calculator and you'll see the settings I mention.


----------



## Nullnvoid

Hugh Grant or Hugh Jackman powered microscope?



Now I have started freezing yeast I have wondered the same thing about how to tell how much is in there in the first place and so how many steps to use. 

I have been using 200ml @ 1.020, 500ml @ 1.040 and 2l @ 1.040 because I read it somewhere.


----------



## Mardoo

Jackman of course. It's not PommyHomeBrewer.


----------



## mofox1

Nullnvoid said:


> I have been using 200ml @ 1.020, 500ml @ 1.040 and 2l @ 1.040 because I read it somewhere.


Depends on what sized vials you are using, how much yeast solids are in the vial, and how old it is.

For a reasonably fresh, 1/2 yeast solids 15ml vial, I reckon that is on the money (anecdotal evidence only). Should also be good for older 50ml vials too.

If there is less yeast solids in the 15's, or they are getting on in age, then I'd recommend using more than one vial.

I've just pitched into 48L of a marzen. Started off at only 10ml solids of 1 year old WLP838. Built up to 11L using 6 steps over about 2 weeks, then 1 week cold crash to decant spent beer so I can pitch just the yeast. 10L of the total was for pitching, the other litre was to put back into the yeast bank.

The lager is currently frothing like mad and making full use of the blow off tube.


----------



## Nullnvoid

mofox1 said:


> Depends on what sized vials you are using, how much yeast solids are in the vial, and how old it is.
> 
> For a reasonably fresh, 1/2 yeast solids 15ml vial, I reckon that is on the money (anecdotal evidence only). Should also be good for older 50ml vials too.
> 
> If there is less yeast solids in the 15's, or they are getting on in age, then I'd recommend using more than one vial.
> 
> I've just pitched into 48L of a marzen. Started off at only 10ml solids of 1 year old WLP838. Built up to 11L using 6 steps over about 2 weeks, then 1 week cold crash to decant spent beer so I can pitch just the yeast. 10L of the total was for pitching, the other litre was to put back into the yeast bank.
> 
> The lager is currently frothing like mad and making full use of the blow off tube.



So much to learn . Love this hobby. 

11 litres? That's one hell of a flask! Where does one by an 11litre flask and then I would love to see the stir plate!


----------



## mofox1

Nullnvoid said:


> So much to learn . Love this hobby.
> 
> 11 litres? That's one hell of a flask! Where does one by an 11litre flask and then I would love to see the stir plate!



LOL. End amount was in multiple flasks. Required significant planning to ensure I had the right flasks at the right time...

I have a 250ml, 500ml, 2 x 1L, 2 x 3L and 1 x 5L flasks + a few other bits and bobs. Missus reckons it a bit of a mad scientist's lab, not inclined to disagree.

*Edit:*
Just for laughs... here is the step up schedule:
21-Jul: pitched 10ml yeast solids -> 300ml(500ml Flask) @ 1.020
23-Jul: 300ml(500ml) -> 500ml(1L) @ 1.038
25-Jul: 500ml(1L) -> 1L(1L) @ 1.038
27-Jul: 1L(1L) -> 2L(3L) for next step & 1L(1L) for vials @ 1.038
29-Jul: 2L(3L) -> 5L(5L) for pitch & 1L(1L) for next step @ 1.038
31-Jul: 1L(1L) -> 2.5L(3L) & 2.5L(3L) @ 1.038, Cold crash 1L(1L) for vials
04-Aug: Cold crash 5L(5L) & 2 x 2.5L(3L)
08-Aug: Packaged 1L(1L) into 7 x 15ml vials ~6ml yeast solids each
12-Aug: Decanted spent starter liquid, vitality starter for 4hrs prior to pitch using wort from cubes, yeast spread equally over 2 x 3L flasks.

All up would have used ~1.5kg of DME... about $9 worth at bulk pricing. Well worth the effort as long you enjoy the process.


----------



## Mardoo

The 11L cubes from DJ_L3thAL are great for doing big step ups. I've gone from 2x 3L grows on the stirplate to an 11L non-stirred ferment to get the amount of yeast necessary for a heavy pitch.


----------



## Midnight Brew

Nullnvoid said:


> I have been using 200ml @ 1.020, 500ml @ 1.040 and 2l @ 1.040 because I read it somewhere.



That is exactly how I step up. Works for both 11L and 22L ferments depending on gravity.


----------



## mofox1

Mardoo said:


> The 11L cubes from DJ_L3thAL are great for doing big step ups. I've gone from 2x 3L grows on the stirplate to an 11L non-stirred ferment to get the amount of yeast necessary for a heavy pitch.


Nice, you get to drink - or more importantly, enjoy - your "starter".


----------



## Mardoo

Yeah, I'm about to do the Westy 12 step up (well, after an IBA ferment) on a hoppy wheat featuring Huëll Melon. I'll rack off the yeast cake for the dry hop. Can't wait!


----------



## Nullnvoid

mofox1 said:


> LOL. End amount was in multiple flasks. Required significant planning to ensure I had the right flasks at the right time...
> 
> I have a 250ml, 500ml, 2 x 1L, 2 x 3L and 1 x 5L flasks + a few other bits and bobs. Missus reckons it a bit of a mad scientist's lab, not inclined to disagree.
> 
> *Edit:*
> Just for laughs... here is the step up schedule:
> 21-Jul: pitched 10ml yeast solids -> 300ml(500ml Flask) @ 1.020
> 23-Jul: 300ml(500ml) -> 500ml(1L) @ 1.038
> 25-Jul: 500ml(1L) -> 1L(1L) @ 1.038
> 27-Jul: 1L(1L) -> 2L(3L) for next step & 1L(1L) for vials @ 1.038
> 29-Jul: 2L(3L) -> 5L(5L) for pitch & 1L(1L) for next step @ 1.038
> 31-Jul: 1L(1L) -> 2.5L(3L) & 2.5L(3L) @ 1.038, Cold crash 1L(1L) for vials
> 04-Aug: Cold crash 5L(5L) & 2 x 2.5L(3L)
> 08-Aug: Packaged 1L(1L) into 7 x 15ml vials ~6ml yeast solids each
> 12-Aug: Decanted spent starter liquid, vitality starter for 4hrs prior to pitch using wort from cubes, yeast spread equally over 2 x 3L flasks.
> 
> All up would have used ~1.5kg of DME... about $9 worth at bulk pricing. Well worth the effort as long you enjoy the process.



Do you chill down between steps or just tip from one into the other?


----------



## Mardoo

I've done and do it both ways, mostly depending on impatience. I'd say not chilling results in the yeast being slightly faster off the mark in the final ferment. In reality though, other than the time it takes to do the rest and chill (72 hours between steps, the way I do it) the difference seems negligible.

If I'm growing up the yeast to freeze it, I do the rest and chill, as (according to the Yeast book) the rest develops glucose reserves, and the chill develops trehalose reserves, both helpful for the post-freeze kickstart.

If I do tip, I check final gravity of the starter steps so I can account for dilution to ensure I stay close to the gravity goal for the follow-on starter. 

Mofo?


----------



## mofox1

Nullnvoid said:


> Do you chill down between steps or just tip from one into the other?


Only after the last step, and not always.

Will almost always decant if pitching into a lighter style beer, OR I'm using a floccing bastard like WLP005 etc that drops without any extra effort. Otherwise, you'll need a couple of days near zero for most yeasts to drop, longer for some non-floccer's.

If have the time to let it drop out, I usually will, as that means I can do a vitality starter prior to pitch.

BTW - Going from a 500ml to 2L, I'll put 200g DME into ~1.7L water, and boil until I'm left with 1.5L (or by weight 1.7kg). I'll then add the 500ml to that to get 2L.


----------



## kaiserben

For the record: after nearly 5 months frozen I've had no problem getting a single vial of WLP029 to take off and can see, from what settled to the bottom after removing from the stir plate, that it most definitely multiplied in my initial 0.3L of 1.020 wort. I'm guessing I now have about 30 ml of solids (would've been only ~12.5 ml or so to start with).


----------



## Yob

So after a very long hiatus from brewing/fermenting, I can see I should get a bit of time to pitch a cube this week so Im spinning up a vial dated 11/14

results to follow, I suspect it was a vial done during my %50 Glycerine ratio as the vial was still mostly liquid (freezer running and holding -20'c) so will give it a spin on the forge and see what happens.


----------



## Nullnvoid

So a silly question but what's the accepted way of thawing the vials?

Sure it's probably been mentioned somewhere in the thread.


----------



## Lord Raja Goomba I

Take out of the freezer.


----------



## Nullnvoid

Lord Raja Goomba I said:


> Take out of the freezer.



And den?


----------



## Danscraftbeer

Then they thaw. I'd think slow and mellow?
I've herd some brewers do it with classical music.


----------



## Tony121

I understand the quicker the better. I whack it in my pocket and take it out & shake every now and then. I also shake the vial whenever I think about it....


----------



## Nullnvoid

So slow and quick . Awesome. Hahahah


----------



## Midnight Brew

Take out of freezer, place near stereo, play Dark Side of the Moon in its entirety, then pitch into a small 1020 starter.


----------



## Mardoo

My kitchen counter seems to have the magic touch.


----------



## Nullnvoid

Thanks everyone. I will slowly but quickly thaw it out with it in my pocket whilst I am sitting on the bench listening to the complete album of dark side of the moon!


----------



## Nullnvoid

Thawed out quicker on the bench than I was expecting.

Alright next stupid question.

I want to use this yeast for a beer but also freeze some. I was going to freeze this batch on the stirplate but leave some and start it again to then pitch into the wort. As the yeast will be healthy can I just start at 500ml @1040 to and go to 2L @1040. Or start smaller again?

Or do something completely different?


----------



## malt junkie

<<< lazy brewer. I make 1L of 1020, start with 200ml, top up over 4days odd till 1L used, then 2L 1040, then 2L 1040. I usually snavel 4-5 vials off this then pitch. Issue is I need to eat more steak.


----------



## Tony121

Nullnvoid said:


> Thawed out quicker on the bench than I was expecting.
> 
> Alright next stupid question.
> 
> I want to use this yeast for a beer but also freeze some. I was going to freeze this batch on the stirplate but leave some and start it again to then pitch into the wort. As the yeast will be healthy can I just start at 500ml @1040 to and go to 2L @1040. Or start smaller again?
> 
> Or do something completely different?


Yes, will be fine to go again


----------



## Yob

so I kind of cocked this one up, largely my own fault I think.

1056 dated 11/2014

flask had been sitting about for a while and I should have given it a stint in the oven.. didnt...

made a 1.040 wort as the first step instead of a 1.020... shit..

after 2 days there wasnt the activity I'd usually expect so I left on the bench to settle out... after a few days it had developed some filming on the top and some bubbles I didnt like the look of, Ive binned that one.. 

The 1.020 wort Ive now produced will have a full tyndallization before adding the yeast 1217 dated 11/2014

starting again... I really want to make sure the first ferment in months hits all the right targets


----------



## Danscraftbeer

Can you pressure cook all plastic vials as like the Whitelabs PET and the Vic Swap give aways etc? For sanitization etc.
I'm just about to test the idea but will they melt or warp at 125c?


----------



## Lord Raja Goomba I

No idea but got some eBay hobbies and they melted. Got some vials which are autoclave able and it's fine to steam heat them.


----------



## Lord Raja Goomba I

*jobbies


----------



## Mardoo

Polypropylene vials are autoclavable. Their melting point is just past 125C, so they squeak by. IIRC PET will not work. The giveaways I can't say. I'd guess they are, but I wasn't there so...


----------



## Danscraftbeer

Its that close then. Bummer. So set the pressure cooker at release option to sanitize the plastic vials at ~100c.


----------



## mofox1

The whitelabs PET blanks do amusing things even in a vegie steamer... think prawn crackers and you aren't too far wrong.

Hate to think what would happen in a pressure cooker.


----------



## kaiserben

kaiserben said:


> For the record: after nearly 5 months frozen I've had no problem getting a single vial of WLP029 to take off and can see, from what settled to the bottom after removing from the stir plate, that it most definitely multiplied in my initial 0.3L of 1.020 wort. I'm guessing I now have about 30 ml of solids (would've been only ~12.5 ml or so to start with).



Following on from the above the rest of the process was simple, as per usual yeast building. 

I'm usually pretty strict about (calculated) pitching rates, but for this lot I just built till the amount of solids looked about what I'd usually have for this type of beer. 

I'm tasting this batch now and it's great.


----------



## Mardoo

Danscraftbeer said:


> Its that close then. Bummer. So set the pressure cooker at release option to sanitize the plastic vials at ~100c.


The polypropylene ones don't deform unless you squeeze them in too tight or rubber band them. Otherwise, since they're just on the edge, they keep their shape.


----------



## DJ_L3ThAL

The ones you recently got in the BB Mardoo have been through a legit dental autoclave (131degC) and came out absolutely perfect no change at all!


----------



## Mardoo

Good to hear! I'll remember that for the future.


----------



## Danscraftbeer

Probably a repeat question but.
Is it best to freeze a liquid yeast from the original vial? I'm thinking yes its the best as to keep the original strain. 
Rather than building it up in a starter and taking from the yeast cake post ferment. I'm thinking just a tad from the fresh vial enough for one vial to freeze then step up the remainder of the vial for the brew day etc.


----------



## Lyrebird_Cycles

DJ_L3ThAL said:


> The ones you recently got in the BB Mardoo have been through a legit dental autoclave (131degC) and came out absolutely perfect no change at all!



The autoclave will run at 121 oC not 131. It is actually 250 oF "metricated". Bloody Septics.


----------



## Lyrebird_Cycles

Danscraftbeer said:


> Can you pressure cook all plastic vials as like the Whitelabs PET and the Vic Swap give aways etc? For sanitization etc.
> I'm just about to test the idea but will they melt or warp at 125c?



Polymers will shift from a glassy state to a rubbery state at a temperature known as the glass transition temperature, Tg. Generally speaking, heating a polymer above its Tg will result in distortion which will "set" once the temperature is lowered so it is sometimes called the Heat Distortion Temperature or HDT.

Low Tg polymers include polyester (PET, recycle code 1, Tg 70 - 80 oC), low- and high- density polythene (LDPE and HDPE, recycle codes 2 and 4, Tgs ~ 90 and ~ 110 oC) and polystyrene (PS, recycle code 6, Tg ~ 100 oC)

None of these can be autoclaved safely.

Higher Tg polymers include polypropylene (PP, recycle code 5, Tg dependant on crystal structure but usually > 120 oC) and polycarbonate , (PC, one of many polymers includes in code 7 "other", Tg around 150 oC).

These can be autoclaved.

Transparent lab gear is often made from acrylic (PMMA) or polymethylpentene (PMP), both are also under code 7 "other". If it's transparent and brittle it's probably one of these, you can tell them apart by colour, PMP usually has a slight yellow cast.

Both polymers have wildly variable Tgs from well below 100 oC to around 150 oC, dependent on structure, so some items are safe and some are not.

BTW code 3 is vinyl or PVC, the heat stability of which depends on the extent of plasticiser used so no generalisation is useful: not much point comparing a down pipe made from UPVC (=Unplasticised PVC) to a piece of flexible vinyl tubing (heavily plasticised, which is why it is flexible).


----------



## hoppy2B

Has anyone mentioned using methylated spirits to sterilise their little tubes and things? I normally just reach for the metho when opening up packets of dry yeast etc.

I haven't been following this thread throughout its entire length, so apologies if it has already been discussed.


----------



## Yob

What brought you back hoppy? A desire to strut your stuff again?

Selling zomes?

There are much better thing to use than methylated spirits...


----------



## Yob

hoppy2B said:


> Has anyone mentioned using methylated spirits to sterilise their little tubes and things? I normally just reach for the metho when opening up packets of dry yeast etc..



https://en.wikipedia.org/wiki/Denatured_alcohol

For wiping a dry yeast, probably not an issue, fucked if I'd put it anywhere near anything coming in contact with my yeast though... but dry yeast isnt this discussion now is it?...


----------



## hoppy2B

Yob said:


> https://en.wikipedia.org/wiki/Denatured_alcohol
> 
> For wiping a dry yeast, probably not an issue, fucked if I'd put it anywhere near anything coming in contact with my yeast though... but dry yeast isnt this discussion now is it?...



Metho evaporates so I wouldn't expect it to cause any problems with yeast as long as you let it evaporate first Yob. 

Nice to see you again too Yob.


----------



## kaiserben

How important do you reckon O2 is for the first step when reviving frozen yeast in ~1.020 wort? 

I ask because my stir plate will probably be being used for something else when I get home tonight, but I really need to un-freeze some lager yeast and get it ready for building up the cell count asap.


----------



## eldertaco

kaiserben said:


> How important do you reckon O2 is for the first step when reviving frozen yeast in ~1.020 wort?
> 
> I ask because my stir plate will probably be being used for something else when I get home tonight, but I really need to un-freeze some lager yeast and get it ready for building up the cell count asap.


I wouldn't worry about it, I've revived from frozen into a jar with 150ml or so of 1.020 and no stir plate quite a few times now.

I just give it a shake every now and then instead, but once or twice would probably be enough I reckon.


----------



## kaiserben

So following on from the above 2 posts I had no problems with minimal O2 in my initial reanimation starter (although there definitely wasn't as much new yeast created as I'd seen with previous yeasts when I did use a stir plate). 

This starter is now on to it's 3rd step at just over 3L and I had krausen sneak out over the lip of my flask and it's now all sticky under the foil (this is a lager yeast). Now I'm going to need to clean the outside and the rim of the flask somehow ...


----------



## Nullnvoid

Need some advice. 

Trying to start some greenbelt which has been in the freezer. 

Started at 200ml at 1.020. Was on the stir plate for 24 hours (probably too long). 

Then 500ml at 1.040. Its currently been 24 hours. No visible action has occurred. Could it be dead? The temperature has been 20-22degrees. 

Should I leave it at that for longer or try to go into 2000ml or something else entirely?

Cheers


----------



## Midnight Brew

Nullnvoid said:


> Need some advice.
> 
> Trying to start some greenbelt which has been in the freezer.
> 
> Started at 200ml at 1.020. Was on the stir plate for 24 hours (probably too long).
> 
> Then 500ml at 1.040. Its currently been 24 hours. No visible action has occurred. Could it be dead? The temperature has been 20-22degrees.
> 
> Should I leave it at that for longer or try to go into 2000ml or something else entirely?
> 
> Cheers



I’d leave it an extra day and assess if the colour of the wort has changed. Currently have the same yeast on 200ml and today into a 1L starter. I’ll keep you updated


----------



## Nullnvoid

Cool. That's what I would have to do. I wouldn't be able to do anything else for 24 hours anyway. 

It's obviously what I got off you. It's been frozen since I got it from you.


----------



## Nullnvoid

So to follow up, leaving it another 24 hours did the trick and it took off. Then stepped up to 2 litres no problems! 

Now I have some vials about to have a sleep and some more back on the stirplate to go into an IPA!


----------



## gezzanet

Just woke up a 1084 Irish ale. Mfg date 21 03 17. Smacked on 22 05 17 and split into 5 x 50ml tubes and frozen. Was cold but not always frozen as drawer freezer was moved and left open once. Yep I have to share it with other food. 
So I thought it might be trashed. 50 ml tube it in tap water to defrost then sprayed with starsan before opening and dumping in 1.020 600ml. 1st day not much action by end of day 2 bubbles showing. 
Stir plate turned off to let it settle out of fridge for 2 hours then poured off 500ml top. 100ml yeast slurry poured into 2500ml 1.030. 25 mm foam in the morning then by 6pm this is what it looks like. Seems hardier than I thought. Think I need to mash an Irish red


----------



## timmyf

Moving house soon so was cleaning out the freezer and came across the frozen yeast bank. I've been enjoying the increasing range of dried yeasts for the last couple years so I'd almost forgotten about it. The oldest vial I had was WLP800 at about 3.5 years old. I decided to spin it up and see how it would go.




Stepped through 200ml of weak wort (~1.020), into 1L of 1.040 and finally 3L of 1.040




End result about a 50ml tightly packed cake of healthy looking yeast.


----------



## Nullnvoid

Stupid question but does anyone make up the starter wort in advance. Ie I have just started the 200ml but make the rest now ready to go for the following steps? 

Too much risk?


----------



## Mardoo

Freeze it if you do, and ideally re-boil. I’ve done it without the freezing step, but for this Nervous Nellie, it was a bit nerve racking up to the point I tasted the beer. Technically boiling can leave some encysted bacteria still viable.


----------



## Nullnvoid

Good enough for me, I'll make it up fresh as the step requires.


----------



## Tony121

This morning I found my beer freezer had been left open the day before and everything in there thawed, including my 40 odd vials of yeast. Everything was still cold but well and truely thawed. I immediately put them in the kitchen freezer whilst dealing with the mess of the other one.

Hoping for some thoughts here on what to expect. Will they be ok or am I going to have to ditch them and start again?


----------



## Tony121

Tony121 said:


> This morning I found my beer freezer had been left open the day before and everything in there thawed, including my 40 odd vials of yeast. Everything was still cold but well and truely thawed. I immediately put them in the kitchen freezer whilst dealing with the mess of the other one.
> 
> Hoping for some thoughts here on what to expect. Will they be ok or am I going to have to ditch them and start again?



Anyone have any thoughts on this?? Dare say the only way to know is to fire them up but was hoping someone had experienced something similar and can offer an opinion.


----------



## timmyf

I'd just relax and not worry too much about it Tony. If they were still cool they should be fine. Not ideal, but fine. 

Mine have always been kept in modern fridge/freezers with defrost cycles that probably push the vials close to thawing. I also had a full fridge blowout about 9 months ago that lead to a full thaw and keeping all the vials at room temp for about two days until I got a new fridge. As you can see in my earlier post-up the page, even after treating my yeast poorly, I still got it to fire up three and a half years later.


----------



## Mardoo

Tony121 said:


> Anyone have any thoughts on this?? Dare say the only way to know is to fire them up but was hoping someone had experienced something similar and can offer an opinion.


Happened to me. All good. I haven't yet had any problems with anything firing up. I did throw out some soup though, but no yeast.


----------



## Tony121

timmyf said:


> I'd just relax and not worry too much about it Tony. If they were still cool they should be fine. Not ideal, but fine.
> 
> Mine have always been kept in modern fridge/freezers with defrost cycles that probably push the vials close to thawing. I also had a full fridge blowout about 9 months ago that lead to a full thaw and keeping all the vials at room temp for about two days until I got a new fridge. As you can see in my earlier post-up the page, even after treating my yeast poorly, I still got it to fire up three and a half years later.





Mardoo said:


> Happened to me. All good. I haven't yet had any problems with anything firing up. I did throw out some soup though, but no yeast.



Thank you both, my blood pressure has come down slightly. 

From now on ice blocks are banned from my beer freezer!


----------



## kcurnow

Attempting to revive some Trappist ale yeast that I have had frozen since Jan 2015. This yeast has so far travelled from Melbourne to Brisbane and may not have been well frozen in the process so it will be interesting to see if it is still viable. 200ml 1020 starter for starters .


----------



## Moad

Nullnvoid said:


> Stupid question but does anyone make up the starter wort in advance. Ie I have just started the 200ml but make the rest now ready to go for the following steps?
> 
> Too much risk?




I went through a stage of canning starter wort but I haven’t been that prepared for a while. You can leave it in the canning jars and have them on hand no dramas. I’m not sure it’s worth the effort though


----------



## MartinOC

Moad said:


> I went through a stage of canning starter wort but I haven’t been that prepared for a while. You can leave it in the canning jars and have them on hand no dramas. I’m not sure it’s worth the effort though


Agreed. Having stuff in liquid form takes up valuable space, when DME can be boiled-up & cooled in an erlenmeyer in the space of 30 mins.


----------



## kcurnow

kcurnow said:


> Attempting to revive some Trappist ale yeast that I have had frozen since Jan 2015. This yeast has so far travelled from Melbourne to Brisbane and may not have been well frozen in the process so it will be interesting to see if it is still viable. 200ml 1020 starter for starters . View attachment 110781
> 
> View attachment 110782



So success!!, after three years of being frozen including a period of being unfrozen when being posted up from Melbourne to Brisbane the WLP500 has fires up after 48 hours in a 200ml 1020 starter. I’m pretty happy with this.


----------



## Midnight Brew

Not a NEIPA but WLP004 frozen (w/15% glycerine) on the 30/11/2014.


----------



## malt junkie

wow, I recon @Yob needs to get his butt in gear and light up a really old one(and possibly brew something for swap), but thats damn impressive MB


----------



## koshari

ok quick question, how much LME in 200ml to make a 1020 starter?


----------



## Midnight Brew

koshari said:


> ok quick question, how much LME in 200ml to make a 1020 starter?



About 10g. I assume LME is light (dried) malt extract not liquid malt extract, as that would be painful to measure.


----------



## koshari

yep i got dried malt extract, thanks, for the quick reply,


----------



## Coalminer

Hi
Have read this thread twice and have some vials frozen but not yet tried one. My question is how do you calculate the viability of the frozen yeast.

First question
when using yeast calculators to build up a pack of yeast for the correct amount for pitching the viability of the smack pack is calculated from the manufacture date, say 1 month old, is 77% and go from there.
My thinking (may be completely wrong) goes like this:
Take a new smack pack, say 1 month old and viability 77% and split into 5 vials of 25ml slurry each = 20x77% = 15.4Billion. Viability when thawed 77% less say 20% loss due to the freezing process = 61% so theoretically 20 x 61% = 12.2Billion - at the date of thawing (use that date in calculator instead of original pack date) use this figure to build starter reqd.

eg Dads Homebrew calculator says 200ml then 1.3L to get 224 Billion for a 1050 23L Ale using the thawing date as viability calculation

Second question
If the above is correct how do you estimate the viability of the following:
Take a smack pack of 1 month old - 77% viable and build a starter to pitch into a brew, but first overbuilding that starter by 100Billion cells. Use whats needed for the pitch and use those 100Billion overbuild cells to split into 5 vials as above.
Can these fresh cells be considered to be 100% as they have just grown, then be considered as 80% viable when thawed (say 20% loss due to the freezing process)


----------



## Midnight Brew

Coalminer, I think you numbers are close to mine. 

Firstly, always make a starter before adding them to vials and the freezer, otherwise you are freezing less ideal yeast while creating a bigger task for the yeast when it comes to reviving. Secondly, the numbers are always going to be ball park estimations. Yeast calculators are great to aid this and will get you numbers that give you a healthy ferment if you have a rough idea on numbers +/- 10%. 

To answer your second question, yes you can assume 100% and 80% viable when thawed (I think it could be closer to 90% but I have no evidence nor microscope).

When I first started freezing, I tinkered around with the numbers a bit to try and keep record how many cells were in each vial, based off the starter size. Now I go with a far easier and what I believe to be more accurate. 3.5-4 billion cells per ml.

I use 15ml vials and by the time the yeast is in there with some water and glycerine it takes up around 40-50% of the vial so 6-7.5ml of yeast. Multiply whats in your vial by 3.5 or 4 and thats how many billion you have. Example: 7.5*4=30 billion cells, minus 20% room for error and thaw = 24 billion cells. 

24 billion cells into a 200ml starter @ 1020 to wake up then whole starter into 1000-1500ml starter @ 1040 usually gets me to where I need for the ales I make. 

Also another handy tip is to keep some old pasta sauce or glass juice bottles. I like to add an extra 5L volume to my brew days when making something around 1040 and use that as starter wort.


----------



## Coalminer

Midnight Brew said:


> Coalminer, I think you numbers are close to mine.
> 
> Firstly, always make a starter before adding them to vials and the freezer, otherwise you are freezing less ideal yeast while creating a bigger task for the yeast when it comes to reviving. Secondly, the numbers are always going to be ball park estimations. Yeast calculators are great to aid this and will get you numbers that give you a healthy ferment if you have a rough idea on numbers +/- 10%.
> 
> To answer your second question, yes you can assume 100% and 80% viable when thawed (I think it could be closer to 90% but I have no evidence nor microscope).
> 
> When I first started freezing, I tinkered around with the numbers a bit to try and keep record how many cells were in each vial, based off the starter size. Now I go with a far easier and what I believe to be more accurate. 3.5-4 billion cells per ml.
> 
> I use 15ml vials and by the time the yeast is in there with some water and glycerine it takes up around 40-50% of the vial so 6-7.5ml of yeast. Multiply whats in your vial by 3.5 or 4 and thats how many billion you have. Example: 7.5*4=30 billion cells, minus 20% room for error and thaw = 24 billion cells.
> 
> 24 billion cells into a 200ml starter @ 1020 to wake up then whole starter into 1000-1500ml starter @ 1040 usually gets me to where I need for the ales I make.
> 
> Also another handy tip is to keep some old pasta sauce or glass juice bottles. I like to add an extra 5L volume to my brew days when making something around 1040 and use that as starter wort.



Yeah thanks MB I think I am the right track. Won't really know until I use some after my upcoming holiday


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## MHB

Braukaiser has some very good reading there is a whole section on storing and reviving yeast, go back to the home page and right at the bottom there are few more bits that really should be linked to the above. There is another section under Yeast Growth Experiments.
Somewhere Kai talked about being able to work out approximately how many cells you have by looking at the change of mass in the starter (if you starter changed 1.020 (20points) or 5oP in 1L you have as oP is %WW, 1L of 1.020 wort weighs 1*1.02=1.020kG, about half of that has been converted to bio-mass (yeast) now divide by the average mass of a yeast cell...That's just from memory go do some digging)
Same bit of info is good for making starters, put the gravity you want in Plato (%WW) if you want 200ml of 1.040 (10oP).
200ml (0.2L) at 1.040 weighs 208g (0.208kg) 10% is extract or 20.8g (0.0208kg).Brewmaths is heally hand
Mark


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## Coalminer

Thanks Mark


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## rossbaker

Hey guys, i finally picked up some vials to give this a go (it's been on the list for ages) . Quick question... Has anyone with a frost free freezer had any luck storing and then refiring these with out doing the whole insulated container thing?


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## portsparky

Old thread I know, but it inspired me to have a go.


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## Quokka42

rossbaker said:


> Hey guys, i finally picked up some vials to give this a go (it's been on the list for ages) . Quick question... Has anyone with a frost free freezer had any luck storing and then refiring these with out doing the whole insulated container thing?



Preferable not to use a frost free anything, as they periodically warm everything up to to melt off the frost.


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## portsparky

For my frost free freezer, I got a soft insulated lunch bag thing from Spotlight .... But noticed these at Bunnings later which looked a bit better than what I got. https://www.bunnings.com.au/esky-9-can-soft-cooler_p3240590

And I'll put some of these soft ice brick in the bags -
https://www.bunnings.com.au/esky-large-ice-cooler-gel-pack_p3240466


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## Tony121

Quokka42 said:


> Preferable not to use a frost free anything, as they periodically warm everything up to to melt off the frost.


Agree it is not the preferred way, though putting vials in a container with ice bricks or something similar does a pretty good job. I have not had an issue this way, still firing up the little troopers after almost 2 years.


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## Tony121

portsparky said:


> Old thread I know, but it inspired me to have a go.View attachment 114533


You won’t regret it, in the top 3 best things I’ve done in regards to brewing. Just make sure you piece together all the good bits out of the thread. Good luck.


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## portsparky

Tony121 said:


> You won’t regret it, in the top 3 best things I’ve done in regards to brewing. Just make sure you piece together all the good bits out of the thread. Good luck.


Thanks Tony. It has taken a lot of reading to try and figure out what people settled on as the best method. I'll just have a go at it and see what happens.

I've attached a couple of other pics. Some were successes, some made my significant other very sad.

Successes included cutting down a rack from proscitech.com.au to fit inside our pressure cooker, and using it to sanitise the 50/50 water/glycerine mix in the tubes. However, I did have to promise first that I'd buy a new one if I ruined the machine. The rack got a little soft bit I think it's fine, and it seemed to work OK.

The yeasties seem happy on the striplate from being on overnight. I'll add another 1 L of wort today.

Failures included me thinking Pyrex would be fine on a cooktop.


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## mofox1

These simple polystyrene boxes work well to insulate against the frost free cycle:





Shame about that Pyrex jug.. while borosilicate does have much lower thermal expansion than ordinary glass, it tends to be less effective with the thicker glass esp when the heat is concentrated in specific locations... That said, I've cracked a flask by trying to dry it off on the cooktop (on low heat). There was just too much heat change too quickly.


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## Tony121

portsparky said:


> Thanks Tony. It has taken a lot of reading to try and figure out what people settled on as the best method. I'll just have a go at it and see what happens.
> 
> I've attached a couple of other pics. Some were successes, some made my significant other very sad.
> 
> Successes included cutting down a rack from proscitech.com.au to fit inside our pressure cooker, and using it to sanitise the 50/50 water/glycerine mix in the tubes. However, I did have to promise first that I'd buy a new one if I ruined the machine. The rack got a little soft bit I think it's fine, and it seemed to work OK.
> 
> The yeasties seem happy on the striplate from being on overnight. I'll add another 1 L of wort today.
> 
> Failures included me thinking Pyrex would be fine on a cooktop.View attachment 114539
> View attachment 114540
> View attachment 114541


Yes a lot of reading, plenty of good info as you get through it though. 

One thing to make sure of is putting the yeasties in the fridge for 24-48 hrs prior to putting to sleep in the freezer - my understanding is that it gives them time to expel the water and soak up the glycerine.


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## mofox1

Tony121 said:


> Yes a lot of reading, plenty of good info as you get through it though.
> 
> One thing to make sure of is putting the yeasties in the fridge for 24-48 hrs prior to putting to sleep in the freezer - my understanding is that it gives them time to expel the water and soak up the glycerine.


^this. Also means if you are using insulated vessel in the freezer, you don't end up heating up your other yeasties by mistake.


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## Kev R

Hi 
Quick questions. Where do you get the little test tubes with lids to freeze yeast?
Are the tubs and lids safe in a pressure cooker?
Is glycerine sterile straight out of the bottle?
Thanks
Kev


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## MartinOC

I got mine from Proscitech in Qld. as part of a bulk buy many years ago.

This is the kind of thing you're after:








10Pcs 50ml Plastic Centrifuge Tube Pipe Vial Lab Test Container With 4 Bot RA | eBay


Find many great new & used options and get the best deals for 10Pcs 50ml Plastic Centrifuge Tube Pipe Vial Lab Test Container With 4 Bot RA at the best online prices at eBay!



www.ebay.com.au





Make sure you get polypropylene ones, which can withstand the temperatures inside a pressure cooker. Get the ones with flat bottoms, as they can stand up on their own.

Nothing is guaranteed sterile with glycerine. You make up a 50/50 mix with water & then pressure-cook the vials filled half-way (lids screwed on lightly).


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## zoigl

Kev R said:


> Hi
> Quick questions. Where do you get the little test tubes with lids to freeze yeast?
> Are the tubs and lids safe in a pressure cooker?
> Is glycerine sterile straight out of the bottle?
> Thanks
> Kev


check my shed clear our ad, I have a huge number of unused t/tubes unused.


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## Moad

reviving ( see what I did there) an old thread. About to spin up some 1968 that's about 5 years old. I can't remember whether to put the glycerine in with the yeast when reviving it. I will decant what I can but from memory a little is good for the yeast


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## Grmblz

Moad said:


> reviving ( see what I did there) an old thread. About to spin up some 1968 that's about 5 years old. I can't remember whether to put the glycerine in with the yeast when reviving it. I will decant what I can but from memory a little is good for the yeast


I use 100ml tubes, my usual process is allow to come up to room temp naturally then chuck the whole lot into 500ml 1035 wort, spin for 3 days crash, decant into 2L 1035 for 3 days, crash decant and pitch, or if in a rush pitch the 2L at "high" adding however much sugar to make up for the 1035, I do use fermaid "O" Not very scientific I know, just seat of the pants stuff. I've seen advice claiming defrosting using warm water to speed up the process is beneficial, have never tried it as what I do works for me.
Good luck with it, I can't remember doing any 5yr old although it's quite possible, been keeping a bank for years but my record keeping is atrocious, I now write the date as well as the strain  fwiw if it hasn't kicked off after 24hrs I ditch it.


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## duncbrewer

I'm just spinning up a Brett salvaged from a keg. Seems to have some foam on the top and active after 4 days with a top up after 3 days. I'm lead to believe that you need to pitch all of a brett starter as it doesn't settle at all well. It's for a 100 brett Saison.


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