# Renewing The Yeast Cultures



## Zwickel (24/3/08)

howdy homebrewers and a happy easter 

The weather is f..ing lousy, around 0C and a little bit snow around here.
o....Im still dreaming about my visit in Western Australia...f....c...k

All of my kegs are filled up with foin Pilseners now, so there is nothing to do at my little brewery.
Today I was racking my latest Pilsener, so I got a lot of fresh yeast.
Then I decided to renew one of my most used yeast strain that Im storing on Agar-Agar.
Basically Im storing a strain on Agar for about one year.

So I started boiling a litre of wort, add some Agar-Agar to it and leave it boiling for a while.
After the boil filled the wort in some vials.
After the Wort cooled down and became stiff, I inoculated each vessel with the fresh harvested yeast.

Now Im the proud owner of more than 25 vessels containing a beautiful Pilsener yeast. That should be enough for one year  

Here a view at my day work.







Id rather go fishing.....on a sunny place like Jurien Bay WA...what Im doing here...

Cheers mates :icon_cheers:


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## blackbock (24/3/08)

How long have you been using this technique Zwickel? and how many generations of yeast have you been using?

BB


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## Zwickel (24/3/08)

Hello BB,

Im applying this method already more than 5 years. Meanwhile I own a pretty little yeast bank.

How to proceed:
First youll need a very fresh yeast, say from a first generation. Store it on Agar as Ive described.
One day, when you need a second batch of yeast vials, just fill one of your first generation vial with fresh wort, keep it for a day and you may inocculate the next batch of vials from that starter. The rest of it you may use for a brew....and so on.

You may do that about 10 times in a row, should be good for the next 10 years.

I know, you wanna ask about mutation and degeneration, no worries.
As long as youre going to use the yeast from Agar to Agar, there is no stress to the yeast and hardly no degeneration and/or mutation, not like in brew.
Copying the yeast from Agar to Agar is very simple and you may do that for the rest of your life.

Cheers :icon_cheers:


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## chiller (24/3/08)

Zwickel said:


> Hello BB,
> 
> Im applying this method already more than 5 years. Meanwhile I own a pretty little yeast bank.
> 
> ...



It is an excellent method. I recently revived a 5 year old "sleeping" 1728 on a slant. It took about 2 days or a bit more to get to a one litre starter. I have not noticed any problems using tis method. When I make my slants I leave them for about 7 days [sealed] without inoculating them and then give them a very close inspection for any growth of any kind. I have only once had a slant that had to be dumped because of contamination. It was very visible to my eye after the seven days.

Zwickel, would you describe your method for reviving the slant up to the starter?

Steve


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## quadbox (24/3/08)

chiller said:


> Zwickel, would you describe your method for reviving the slant up to the starter?



I'm also quite interested in this. Planning on doing up my first slants next week (some wlp-023 and some wlp-001), that's the only part of the process I'm not entirely clear on. Have seen a couple of different ways suggested.

As an aside, someone on another forum pointed out these guys (no affiliation) who sell 50 packs of screw-top test tubes relatively cheap, see down the page


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## pint of lager (24/3/08)

Zwickle, try adding a bit less agar/malt solution to the tubes and laying them down a bit further. This will give a greater surface area for the yeasties to do their thing.

Then, as chiller suggested, leave them for a week to allow for any infected tubes to show up before innoculating them.

Otherwise, a fine day's work.


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## ausdb (24/3/08)

Zwickel said:


> howdy homebrewers and a happy easter
> 
> The weather is f..ing lousy, around 0C and a little bit snow around here.
> o....Im still dreaming about my visit in Western Australia...f....c...k
> Cheers mates :icon_cheers:


Hi Zwickel that weather is a bit different to Kooks house on Australia day, maybe we should send the 4 kettles your place to warm you up a bit!!

Thanks for the info it's something I have been meaning to try for a long time, you make it seem quite simple.


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## chiller (24/3/08)

Here is my method of reviving a slant.

Method [mine] Sterile cooled starter -- about 5 or 10 ml into the slant tube. Shake and re seal. Over the next 24 hours gently ease off the cap with it right next to your ear. When you hear a phwwwt!! [  gas escaping noise] that is the time you have enough to add to about 50ml of starter. Leave for 24 hours and then transfer to 240 ml of starter after another 24 hours step up to about 500 - 1000 ml.

Steve


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## Pumpy (24/3/08)

I wish I could do all that agar stuff to save yeast!! is it that hard !!!would save a fortune 


Pumpy


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## chiller (24/3/08)

Pumpy said:


> I wish I could do all that agar stuff to save yeast!! is it that hard !!!would save a fortune
> 
> 
> Pumpy



There are other methods -- but -- no matter which method you choose, sanitation is paramount. I use both slants and storage under distilled water. The distilled water method has been very good for me. Slants are a bit like the Rolls Royce of yeast ranches. The little yeasties seem to go to sleep and the moment you re-introduce some food they take off where they once were. I only brew Ales so cannot comment if "girlie" lager yeasts are as robust. I would imagine they are.

My Belgian yeast collection is under water with a backup slant.


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## Pumpy (24/3/08)

Chiller is there any step by step posts on this process ?????



pumpy :unsure:


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## Pumpy (24/3/08)

wally said:


> Pumpy, have a look at this link.
> 
> 
> Wally




Wow Nothing short of Excellent thanks Wally 

pumpy


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## quadbox (24/3/08)

Two others in my bookmark tree: here and here


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## Zwickel (24/3/08)

pint of lager said:


> Zwickle, try adding a bit less agar/malt solution to the tubes and laying them down a bit further. This will give a greater surface area for the yeasties to do their thing.


yeah, thats right, but this way it works fine too:





If you lay it down further, youre in a danger the liquid flows out and if the vessel is contaminated outside, you have to threw it away.



chiller said:


> Zwickel, would you describe your method for reviving the slant up to the starter?



okey-dokey, I didnt know thats interesting to you.

After one week of inoculation (keep it for the first week at ambient/room temp), Im storing the slants in my converted freezer, around 2C.
Whenever I need a starter, Im taking one of it, rewarm it to around 20C. Meanwhile I have taken 1litre of frozen wort and rewarm the wort to 20 too.
Fill in some wort into the slant, close the lid and wait. Keep the rest of the wort in the fridge. 
After a day, depends on how old the slant already is, sometimes it needs two days, you may see very fine little bubbles on the surface.
Thats the sign the fermentation has begun.
Now Im taking a larger vessel, say an Erlenmeyer with a size of one litre. Transfer the content of the slant into the Erlenmeyer add about 100ml wort (rewarmed in the microwave to 20C) put in a magnetic fish and put it on the magnetic stirrer, start stirring.

After another day you may see now a huge foam head in the vessel, now you transfer the content into a larger vessel, Im taking 5l bottles, add the rest of the wort, stirr well and start airaiting it and start cooling down slowly. For cooling I put the whole equipment into my fermentation fridge.

here another pic:





Then after another day add some more wort to it, depends on how much yeast youll need.
Meanwhile the temp has reached about 10to 12C.

Leave th wort over night to settle and decant next day, fill in more fresh wort.

In the end it looks like that:






thats all. (apologies for my poor english)

Cheers


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## quadbox (27/3/08)

I'm going to be putting together my first slant collection next week, should be fun. Have picked up fifty of those v-bottom free standing test tube thingies with the screw-tops, will be doing up about twenty WLP-023 (aka wyeast 1275 iirc), twenty WLP-001 (wyeast 1056) and a few wlp-001's, mostly because I have some around and figure I'll do some side-by-side's with the burton ale.

That should last me a while. Will save me quite a lot all told, until fairly recently was paying through the teeth for sachets of saf for every brew. at $4 or so a brew, adds up quick


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## Zwickel (27/3/08)

that little bastards have been very hungry and over active; this morning I found one of the slants bursted, completely destroyed from overpressure.

Guess I kept it a little bit too warm, around 24C.

So carefully I opened each vessel just a little and immediately closed it again, so there wouldnt be any pressure left inside.

It never happened to me before.

Im gonna punish them, put em in the cold now.  

Cheers :icon_cheers:


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## Kleiny (27/3/08)

Thanks guys
this has made me want too have ago at slants i always thought they were a bit too hard
but the links and chat on this forum have made it a lot clearer

cheers


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## Franko (27/3/08)

Im going to give this a go.
Pumpy if your interested give us a call and we'll get off to a start

Franko


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## alexbrand (28/3/08)

Franko said:


> Im going to give this a go.



It's really an excellent method for yeast farming. I do it, too: Picture

Alex


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## Sprungmonkey (28/3/08)

Hi all. Quick question when making a starter from cultures in the slants people are stepping up from 50 to 250 to 500 to 1500mL. Is there any real point in stepping up to 1500ml. I don't really understand why so much is made up for the starter? Could you simply add to the fermentor at 250ml?


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## Zwickel (28/3/08)

yeah, youre right, if you are using top fermenting yeasts, you dont need that much.

The most of my beers are Lager or Pilsener beers. As you know, when using bottom fermenting yeasts, you need much more of it than using top fermenting yeast. Also Im doing 60l batches, so I have to use about 1% of the batch size stiff yeast, thats about 600ml stiff yeast.

For top fermenting beers like Ales, youd need less than the half of it, say 0.3% of the batch size.

Cheers


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## Zwickel (29/3/08)

Just to complete the procedure of yeast farming.

how to inoculate:

Im taking one of that little thingy:







sterilized over hot steam, to lengthen it, you my use a skewer as well, dip it into the yeast slurry and then just roll it over the agar.

Dont fill yeast into the slant! just touch the agar with a contaminated cotton-wool swap.

At first you shouldnt see any yeast on the surface of the agar. After a couple of days you will notice some thickness on the surface, later it will turne grey.


Thats all.

Cheers :icon_cheers:


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## BenH (30/3/08)

Thank you Zwickel. You've demystified the process - I'm going to give it a go now!


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## Sprungmonkey (3/4/08)

Zwickel - just wondering how much agar agar to water to DME you use for making the slants? Also do you add other nutrients?


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## ausdb (3/4/08)

Zwickel said:


> After another day you may see now a huge foam head in the vessel, now you transfer the content into a larger vessel, Im taking 5l bottles, add the rest of the wort, stirr well and start airaiting it and start cooling down slowly. For cooling I put the whole equipment into my fermentation fridge.
> 
> here another pic:
> 
> ...


Zwickel can you please post a picture of the stir bar that you are using in the 5l bottle, I know the bottom of the bottle is convex and I was wondering how well the stir bar stays centered or do you have problems with it flying off?


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## Zwickel (3/4/08)

Sprungmonkey said:


> Zwickel - just wondering how much agar agar to water to DME you use for making the slants? Also do you add other nutrients?


howdy homebrewers,

I dont use any DME although it would be right using it.

Im producing about 70 to 75 liters of wort every brew Im doing, but cant ferment more than 60litres of it.

The overplus Im gonna freeze in one liter portions. So whenever I need fresh wort for starters, Im taking one part of it
and no, I do not add any other nutrition.

Im using around 5g of agar for 1 litre of wort. The agar you may get at any Asian Food Shop, costs really peanuts.
Before youre going to add the agar to the wort, it would be better if youre dissolving the agar powder in cold water, it avoids clumping.

Fortunately I own a freezer/fridge combination exclusively for my brewing stuff. The freezer is used to store hops and wort and the fridge for fermenting.

@ausdb, no worries, the stir bar centers immediately when you put it into the bottle. It works very well.

Later at the afternoon I will post you some pics of it, right now Im sitting at my workplace, remember, were about 8 hours behind you.

Cheers :icon_cheers:


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## AndrewQLD (3/4/08)

> Later at the afternoon I will post you some pics of it, right now Im sitting at my workplace, remember, were about 8 hours behind you.



That sucks Zwickel, I bet you wish you lived here in Australia, that way you would be Knocking off work right about now and tucking into a beer.

Never mind, I'll have a Weizen for you now :lol: 

Cheers
Andrew


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## Zwickel (3/4/08)

AndrewQLD said:


> That sucks Zwickel, I bet you wish you lived here in Australia, that way you would be Knocking off work right about now and tucking into a beer.



Mate, you dont know how true that is 



> Never mind, I'll have a Weizen for you now :lol:



I hope, next holiday of mine we can have beer together next eachother.

Cheers :icon_cheers:


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## ausdb (3/4/08)

Zwickel said:


> @ausdb, no worries, the stir bar centers immediately when you put it into the bottle. It works very well.
> 
> Later at the afternoon I will post you some pics of it, right now Im sitting at my workplace, remember, were about 8 hours behind you.


Thanks very much, I have 2l and 3l erlenmeyer flasks and have trouble keeping the stir bar centred even though they are flat bottomed when there is a large yeast load. I was thinking of using a 5l bottle like yours but thought I would have trouble keeping the bar centred because of the domed base.


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## Zwickel (3/4/08)

ausdb said:


> Thanks very much, I have 2l and 3l erlenmeyer flasks and have trouble keeping the stir bar centred even though they are flat bottomed when there is a large yeast load. I was thinking of using a 5l bottle like yours but thought I would have trouble keeping the bar centred because of the domed base.


yeah, Ive used Erlenmeyer flasks too, but in my Opinion, the bottles are much more handy.

Your trouble keeping the stir bar centered could be even because of the flat bottom.
Maybe the stir bar spins much easier if the bottom is domed, due to the friction, just a thought.

Anyway, I will post some pics Ive already taken.

Cheers :icon_cheers:


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## ausdb (3/4/08)

Zwickel said:


> yeah, Ive used Erlenmeyer flasks too, but in my Opinion, the bottles are much more handy.
> 
> Your trouble keeping the stir bar centered could be even because of the flat bottom.
> Maybe the stir bar spins much easier if the bottom is domed, due to the friction, just a thought.


I will try it as I went to buy a new stir bar with the ring around it yesterday but the lab supply place had sold out. One thing that I do like about erlenmeyer flasks is being able to put them on the stove and boil the starter wort and stir bar in the same flask that I will be culturing my yeast in as I use DME for my starters. It means I can cool it in the sink then pitch the yeast in and not worry about sanitising more containers or unnecessary transfers, but a 5l demijohn like you use costs only $17 whilst a 5l erlenmeyer costs over $50.


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## Zwickel (4/4/08)

ausdb said:


> I will try it as I went to buy a new stir bar with the ring around it yesterday but the lab supply place had sold out. One thing that I do like about erlenmeyer flasks is being able to put them on the stove and boil the starter wort and stir bar in the same flask that I will be culturing my yeast in as I use DME for my starters. It means I can cool it in the sink then pitch the yeast in and not worry about sanitising more containers or unnecessary transfers, but a 5l demijohn like you use costs only $17 whilst a 5l erlenmeyer costs over $50.


Thats fine mate, of course an Erlenmeyer flask looks more professional than a bottle of "Hobos Heaven" 
but for myself I found the bottles more handy.

oooops....A$17 for that bottle? We pay 4.98 Euro for 5l of red or white wine in that bottles, just the half of it  

thats my collection of stir bars, the upper one (on top of the pic) is the one Im using most:








here you can see a test run with water and airation.
I dont use that plastic diffusor anymore, Im now using a sintered brass diffusor as you can see in the pic above:






and here something you may lough at:







that looks quite funny, but its nice to see how much CO2 is produced as the balloon gets big and bigger.

Cheers :icon_chickcheers:


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## kabooby (18/4/08)

From the info in this thread I have recently started to store my yeast on Agar.

I used to just store the yeast in stubbies but this was starting to take up to much fridge space.

I have made up 24 slants and just innoculated 2 with some Burton ale yeast WLP023, so well see how they go.

What is the main purpose of the Agar? Couldn't you just make up a malt mixture and store it just the same? The obvious difference is that the Agar makes the malt solution solid, but what is the advantage of this?

Kabooby


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## matti (18/4/08)

Agar is food for cell!?

correct me if I am wrong.


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## Franko (18/4/08)

Kabooby 
what did you use for your slants - tube wise


Franko


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## Sammus (18/4/08)

Here's another useful link in culturing - maybe a little more tech and finicky, requiring that everything is sterilized properly (ie autoclaved/pressure cooked).

Also, I noticed Zwickel said to dissolve the Agar in cold water. Maybe disperse it through? One of the properties of that Agar stuff is that it won't dissolve until the liquid is nearly boiling.

Also, boiling stir bars is a bad idea - they loose their magnetism when heated up. I lay out some glad wrap, spray with some iodophor and keep it wrapped up, then just drop it in when I need it. The glad wrap is good because you can handle the stir bar through the film without touching it directly.


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## Zwickel (18/4/08)

Sammus said:


> Also, I noticed Zwickel said to dissolve the Agar in cold water. Maybe disperse it through? One of the properties of that Agar stuff is that it won't dissolve until the liquid is nearly boiling.


have you ever done it? you can see the difference very well. Put it in hot wort and youll get clumps, hard to dissolve. No problem in cold wort.



> Also, boiling stir bars is a bad idea - they loose their magnetism when heated up. I lay out some glad wrap, spray with some iodophor and keep it wrapped up, then just drop it in when I need it. The glad wrap is good because you can handle the stir bar through the film without touching it directly.



the stir bars are made to get autoclaved. Otherwise you could use a stirbar only once.
After use Im sterilizing the bars in a steam pressure cooker.
We have stir bars in our laboratory they get used since more than 10 years and steady autoclaved, they havent lost theire magnetism, not at all.

anyway, its not a good idea to stir the hot wort with a magnetic stirrer, thats simply unnecessary, just boiling is enough.

Cheers :icon_cheers:


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## kabooby (18/4/08)

matti said:


> Agar is food for cell!?
> 
> correct me if I am wrong.



I thought the malt was the food for the yeast, could be wrong though.



Franko said:


> Kabooby
> what did you use for your slants - tube wise
> 
> 
> Franko



Hi Frank,

I am using the LP9025 UL containers from here

Got 100 of them :blink: 


I will try dissolving the Agar in cold water next time. It did clump in the hot water.

So is the Agar food for the yeast as well as the malt?

Kabooby :icon_cheers:


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## Zwickel (18/4/08)

the food for the yeast is in the wort, mainly the sugar thats in.
Agar is nothing for the yeast, is basically colourless, odorless and flavorless, just makes the wort stiff.

The trick is, the yeast spreads over the surface of the agar, dissipating all the nutrients it can get and falls asleep.
In a solution, the yeast would be affected by the alcohol and autolysis, not so on the surface of an agar bottom.

One day when you fill in a little bit wort into the slant, only the yeast cells alive will start to regenerate and get into the wort.
All other crap like dead yeast cells will remain in the agar. So after you have transferred the initial wort to a bigger vessel, you have only living cells extracted (theoretically)

Cheers :icon_cheers:


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## mfdes (18/4/08)

Hi Zwickel,

I notice in your post that you had the wort on a stirrer and ALSO were adding aeration. Have you found the added aeration to make a significant difference?
I get very good growth just on the stirrer, but was wondering whether adding extra air is worth the contamination potential.

MFS.


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## Zwickel (18/4/08)

mfdes said:


> I notice in your post that you had the wort on a stirrer and ALSO were adding aeration. Have you found the added aeration to make a significant difference?
> I get very good growth just on the stirrer, but was wondering whether adding extra air is worth the contamination potential.



it depends on the time I have and what kind of beer Im gonna brew.
Of course, there is a significant difference in growth.
Just from feeling I would say that the yeast will grow 5 to 10 times faster if the starter gets additional airated.

For Pilseners you cant get a starter big enough for 60l of wort by stirring only, within one week. As you may have seen in my pictures, I need at least 500ml stiff yeast.
Thats completely different if you are using top fermenting yeasts, such as for an Ale or Hefe Weizen. For that, stirring only would be far enough. 



Cheers

btw. I never had any infection


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## kabooby (18/4/08)

Thanks Zwickel, Makes sense

Kabooby :icon_cheers:


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## andreic (27/4/08)

This is an interesting thread. I'm thinking of giving the slants a go. One question though...

How long do the slants last in the fridge?

Some of the links posted in this thread suggest 3 months! Then you have to "renew" the yeast. But some of the posts in the thread suggest up to a year!

Up until now I've been storing my liquid yeasts in stubbies after growing a starter from the original whitelabs tube or wyeast pack. This takes up a lot of room. I also reckon some of my older yeasts were not quite in "optimum" state when revived. My last hefe, whilst ok, lacked somewhat in the aroma and flavour department when compared to the previous beer prepared exactly the same way. I'm looking at slants as a way to keep yeast in better condition for longer... but how long?

cheers


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## Hogan (27/4/08)

pint of lager said:


> Zwickle, try adding a bit less agar/malt solution to the tubes and laying them down a bit further. This will give a greater surface area for the yeasties to do their thing.




Had some deep and meaningful discussions on yeast culturing yesterday with Franko and Kabooby. (The more pubs we attended the more D & M it became). Discussed slants and what we would use to grow the yeast in - tubes etc. After clearing my head this morning I considered that the reason the tubes are slanted was to get as large a surface area for the yeast cells to cling to the agar. I found a small jar which had apparently been used for jam. Its volume is 30ml and it stands 4cm high with a screw top metal lid. These would give a much larger surface area than a tube and could be sterilised as effectively. I have attached a pic. Have looked at a number of sites for jars and Lab equipment but none seem to have this size. Any ideas where I might be able to procure them. They appear to be a type that may be used on airlines or for samples of food etc. Any suggestions would be appreciated.

Cheers, Hoges.


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## Duff (27/4/08)

Agar is also cooled on a slope in laboratory uses so that any moisture also collects at the lowest point and not on the agar itself. If agar is mixed with any bacterial inhibitor and moisture gets on the surface, the inhibitor is diluted and contamination occurs. 

Same principle for your yeast slants, less moisture, less contamination on the surface of your agar.


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## Zwickel (27/4/08)

andreic said:


> How long do the slants last in the fridge?


gday mates,

the professionals suggest not to exceed 3 month. Thats for professional brewing like in the industry.

I have used and successfully revived slants even after 2 years. I cant say anything about the quality of that yeasts, but I can say that the beer was made from didnt showed any flaws.

@Hogan, of course you may use any type of glass that hermetically can be closed. At my start on yeast farming Iv used mainly glasses where baby food had been in. It worked well all the time. Only, because my yeast collection became larger, I had to switch to smaller vessels.

@Duff, what you said is right, its recommended to keep the agar as dry as possible. Never put in a drop of liqid yeast for inocculation. That would be far too much moisture. Just roll a cotton wool swap over the surface, thats enough.

In case youd get too much moisture in your vessel, the yeast may get under the agar and its CO2 production leaves the agar floating.

Cheers :icon_cheers:


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## kabooby (27/4/08)

Zwickel said:


> In case youd get too much moisture in your vessel, the yeast may get under the agar and its CO2 production leaves the agar floating.



I have had this happened now to one of my slants. I haven't used them yet so I am assuming they will still be ok.

Kabooby


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## Zwickel (27/4/08)

kabooby said:


> I have had this happened now to one of my slants. I haven't used them yet so I am assuming they will still be ok.
> 
> Kabooby


yeah, when I started to work with agar, I was always concerned to get too less yeast in, so I put in far too much yeast and moisture. All of my agars started floating in the glasses. That time I didnt know why the agar got a lift up, only later I realised, that its the Co2 that gives it a lift.

Anyway, as long as you dont get any strange germs in, it doesnt matter, it just doesnt look nice. I never had any infected agar.

Cheers :icon_cheers:


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## Leigh (27/4/08)

ausdb said:


> Thanks very much, I have 2l and 3l erlenmeyer flasks and have trouble keeping the stir bar centred even though they are flat bottomed when there is a large yeast load. I was thinking of using a 5l bottle like yours but thought I would have trouble keeping the bar centred because of the domed base.



From my lab days, the size of the magnet in the magnetic stirrer, the size of the stir bar, the speed of the stirrer and the viscosity of the solution all affect the stability of the stir bar...if you are not getting the stir bar to centre, try a smaller stir bar and/or start stirring slow and gradually increae the speed...even then, sometimes you can not get up to the maximum speed of the stirrer.

Flat bottom glassware assisted the stability...round bottom flasks needed quite small stir bars to stay stirring centrally.


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## Sprungmonkey (28/4/08)

These are my first slants


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## kabooby (28/4/08)

Look good Sprungmonkey :icon_cheers:


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## alexbrand (28/4/08)

Sprungmonkey said:


> These are my first slants



Reminds me of a urine test...  *g*

Just kidding, looks great. How did you prepare the slants?
To prevent infections I got used to put the whole filled tubes at the end into a pressure cooker for some minutes. So i don't have to work that clean in all the different steps before. Saves time and protects my nerves. If the tubes are deliverd sterile there's no need to do so...

Greetings,

Alex


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## Sprungmonkey (28/4/08)

alexbrand said:


> Reminds me of a urine test...  *g*
> 
> Just kidding, looks great. How did you prepare the slants?
> To prevent infections I got used to put the whole filled tubes at the end into a pressure cooker for some minutes. So i don't have to work that clean in all the different steps before. Saves time and protects my nerves. If the tubes are deliverd sterile there's no need to do so...
> ...



Made up media (wort and agar agar) in a 1L Schott bottle, Autoclaved (15mins @ 121C), poured into centrifuge tubes and let set. Luckly I have access to an autoclave.

Cheers
Dave


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## Hogan (28/4/08)

Sprungmonkey said:


> These are my first slants




These are the same size Franko was thinking of getting. 50ml is a good size and there seems to be plenty of surface area on the slant. Thanks for the pic Dave.


Cheers, Hoges.


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## Sprungmonkey (30/4/08)

One other thing. I always have a flame and metho handy for sterilising inoculating loop and after opening jars etc. It just good practise to quickly flame lids and lips of containers when pouring transfering etc. 

Just thought I would add that.


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## maltedhopalong (30/4/08)

I have a little question...

Will a yeast keep better (healthier, more viable cells etc.) in a slant or in pure water in the fridge?

The reason I ask is that it would take less space/time/money to just keep one jar of yeast in the fridge in pure water and when you create the starter, you make one bigger than you need, pitch half and then dilute the other half with pure water and put it back in your original jar...

Suppose there's a good reason why this won't work tho


----------



## alexbrand (30/4/08)

Sprungmonkey said:


> Made up media (wort and agar agar) in a 1L Schott bottle, Autoclaved (15mins @ 121C), poured into centrifuge tubes and let set. Luckly I have access to an autoclave.



Your're really lucky!  But... aren't those sample tubes autoclavable? So I would sterilize theready made tubes. This is even more secure and consumes the same time, I think?




maltedhopalong said:


> Will a yeast keep better (healthier, more viable cells etc.) in a slant or in pure water in the fridge?



On a slant! In pure water the yeast suffers from famine.  On a slant the cell will find food in the begining. The cell consumes it and propagates. But soon it'll "fall asleep" as long as no nutrients are available. If you "start" the slant later with frsh worth. The cells wake up and get busy... 

Greetings,

Alex


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## maltedhopalong (30/4/08)

> the yeast suffers from famine



Suffers how? Does it die or just go to sleep? Considering the effort you go to to make a starter from an agar slant, I'm thinking if it only goes to sleep, it's easier to just keep a jar of washed yeast in the fridge and pitch the whole jar to start a new brew. Then collect the trub, wash and keep THAT in the fridge. Less (or same) work, less cost and presuming you don't need to do two brews with the same yeast at the same time, endless supply.

Sorry to hijack the thread with more low-level questions.


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## Zwickel (30/4/08)

maltedhopalong, the answer of your question Ive given already in post#41 in this thread.

in a liqid the yeast will start to degenerate and suffers from autolysis.
Later, if youre going to do a starter from that kind of yeast, youll get all that shit into your brew.

Beside, one cannot wash the yeast to separate the dead cells from living ones, they are at the same size and weight.

Cheers :icon_cheers:


P.S. if youre going to brew fortnightly, then it would be fine to keep the yeast just in the fridge. On agar its more a long term storing method.


----------



## maltedhopalong (30/4/08)

cheers Zwickel, sorry mate.

I did read your post but obviously didn't understand it 100% as I've never done a slant myself (specifically about how washing doesn't have the same effect).

Like I said, sorry about the noob angle, but appreciate your humouring me anyway.


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## Zwickel (30/4/08)

no worries mate,

thats what this forum is created for, just to spread the knowledge.

Can you imagine, that a certain amount of harvested yeast containes not only living yeast cells?
It is simply not possible by washing the yeast to separate the dead or degenerated yeast cells from the good and healthy one, nor is it possible to separate floating debris that is at the same size and weight like the yeast cells. The only thing you can do whilst washing is to dilute the liquid and lower its alcohol content.

So in the end youll get all that unwanted stuff into your starter and later into your beer.
If there are any strange germs in your yeast, they will multiply over the time.

On an agar bottom, if youre going to revive one, youll only extract living yeast cells, all that unwanted stuff remains in the agar.

Cheers


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## alexbrand (30/4/08)

maltedhopalong said:


> Suffers how? Does it die or just go to sleep? Considering the effort you go to to make a starter from an agar slant, I'm thinking if it only goes to sleep, it's easier to just keep a jar of washed yeast in the fridge and pitch the whole jar to start a new brew. Then collect the trub, wash and keep THAT in the fridge. Less (or same) work, less cost and presuming you don't need to do two brews with the same yeast at the same time, endless supply.



Zwickel already answered the question, nothing to add. 

I have no idea who postulated the theory, a home brewer can wash yeast. Well, I mean, of course one _can_ wash yeast. But with rather no effect. It might be possible to seperate cells from other rubbish. But not cells from cells. Doesn't matter if it's yeast, bacteria, virus, prions or what ever. 

Happy yeast farming!

Alex


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## mikelinz (1/5/08)

One of the links on slanting said to let the medium dry out as moisture in the slant can lead to mold etc. So How do I dry it out if it is sealed.


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## mikelinz (1/5/08)

Is there a list where you can post yeast types/slants you have available for swap etc

rgds mike


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## Zwickel (1/5/08)

mikelinz said:


> One of the links on slanting said to let the medium dry out as moisture in the slant can lead to mold etc. So How do I dry it out if it is sealed.


how mould can grow if everything is steril?

If you got mould spores into the slants, the mold will grow whether the agar is wet or dry, other hand, If everything is steril, no mould can grow.

Since Im doing yeast farming I had almost every form of agar, from very wet to very dry, neither of them ever had any mould in.

Anyway its recommended to keep the agar as dry as possible, one procedure could be:

sterilize the vessels in a baking oven (without the cap). Fill in the hot agar at around 100C and leave it in the baking oven. Slowly cool down and do a rest at around 40C. after a half an hour open the baking oven and cap the vessels. voil thats it.

Cheers :icon_cheers:


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## Zwickel (1/5/08)

mikelinz said:


> Is there a list where you can post yeast types/slants you have available for swap etc
> 
> rgds mike


Mike, much to my regret, Im not living in Australia, otherwise Id like to give away many of my yeast strains. 
But as more and more homebrewers get into yeastfarming, maybe one can organize a swap meet or do an exchange via mail.

Cheers


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## Sprungmonkey (1/5/08)

Zwickel - Im in


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## quadbox (6/5/08)

Here's my first effort, so far:



















Those lids are only on loosely atm to let the moisture dry out a bit before I seal them. Planning to leave them for a week or so to check for infection, then inocculate 10 with wlp-001, 10 with wlp-023, and five with wlp-002


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## Duff (6/5/08)

Well done, they look good. No need to leave for a week to check infections, 2 - 3 days at room temperatures will suffice.

Cheers.


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## remi (6/5/08)

Looks the goods....fun isn't it?

remi


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## Sprungmonkey (6/5/08)

I used one of my first slants yesterday (after preparing a starter friday last week). Used a whitelabs mexican lager. Also made some San fransico lager ones as well. The cervasa is bubbling like crazy so it appears to have worked. Looking forward to the end results.


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## Sprungmonkey (6/5/08)

If the final results are all good I don't think Ill ever used dried yeast again


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## quadbox (6/5/08)

Sprungmonkey said:


> If the final results are all good I don't think Ill ever used dried yeast again



Well I mean just from a price point of view, you're looking at like 80c worth of dme for a 1L starter, perhaps another 80c worth of slant, vs $4 or more per sachet of saf. That works out to be a fair saving over time imo, especially given you can keep cultures of a wide variety of yeast


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## Sprungmonkey (6/5/08)

I didn't use as stirrer at all with the starter but stirred it evey time I walked past. I think Ill get one of those next.


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## Zwickel (6/5/08)

wow, that looks very nice  

well done mate. :icon_cheers:


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## Sprungmonkey (6/5/08)

Zwickel said:


> wow, that looks very nice
> 
> well done mate. :icon_cheers:




Thanks for your help


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## tynian (7/5/08)

Thanks to this thread, I made my first slants on the weekend. It just seemed all to easy.

I am going to do some Wyeast 2001 (Urquell) this weekend. Then I hope to move some of my 3944 to slants for safe keeping.

It would be great if there are any other peoples interested in doing swaps etc.

Rob.


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## Steve Lacey (7/5/08)

1) Why are all you people wasting a whole slant each time you brew? Just use a sterilized (flame or alcohol) loop or needle or whatever to pull out a blob of yeast from the slant and step up a starter going: 20 mL ... 200 mL .... 2000 mL (or less). And your slant will be good to use again tomorrow, next week or next year (conditional to the answer in part 2). The photos with rows and rows of slants seems unnecessary unless the user is about to culture up a dozen or so different strains. You should never need more than three slants of any variety, and even then two of them are reserves.

2)


> Some of the links posted in this thread suggest 3 months! Then you have to "renew" the yeast. But some of the posts in the thread suggest up to a year!



The thing about yeast is that it is a living organism and there are all kinds of variations that affect the answer to "how long can I keep a slant?" Differences between strains, differences in the quality of the medium, differences in your level of sterility. So mentions of "three months" means in most cases you should be very safe for up to three months. After that, you could be fine for another 12 months or 2 years or more if you look after the thing. But the _certainty _of it all being good just gradually declines, so at some point to get yourself back into safe territory you make some new slants and innoculate them with a tiny dab of yeast from one of the old slants, provided the yeast still looks clean.


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## ~MikE (7/5/08)

tynian said:


> Thanks to this thread, I made my first slants on the weekend. It just seemed all to easy.
> 
> I am going to do some Wyeast 2001 (Urquell) this weekend. Then I hope to move some of my 3944 to slants for safe keeping.
> 
> ...



should we perhaps set up a AHB wiki page where people post up their stocks of yeats. then we could just PM trade via mail etc 

but yeah, i'm definitly up for it.


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## AndrewQLD (7/5/08)

Steve Lacey said:


> 1) Why are all you people wasting a whole slant each time you brew? Just use a sterilized (flame or alcohol) loop or needle or whatever to pull out a blob of yeast from the slant and step up a starter going: 20 mL ... 200 mL .... 2000 mL (or less). And your slant will be good to use again tomorrow, next week or next year (conditional to the answer in part 2). The photos with rows and rows of slants seems unnecessary unless the user is about to culture up a dozen or so different strains. You should never need more than three slants of any variety, and even then two of them are reserves.



Steve, I think the idea behind this thread is to be able to use the whole vial as a starter, Zwickel posted his method early in the thread and Chiller added a more complete description which included his system of adding boiled cooled wort to the inocculated vial, shaking and leaving overnight to create a mini starter of sorts. This is then pitched into a larger sized starter ect.

Andrew


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## Zwickel (7/5/08)

gday mates,

yeah, there are many ways to treat the slants.

My first intension was to show our fellow brewers how easy yeastfarming could be. Just to encourage the brewers, they have no glue about biological stuff, to get into.
My idea was to use only equipment everybody has at home, not to make it too complicated.
Im very glad to see, many of you have taken the bait 

After getting the foot into the door, one may elaborate individually theire own methode.

Cheers mates :icon_cheers:


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## Steve Lacey (7/5/08)

AndrewQLD said:


> Steve, I think the idea behind this thread is to be able to use the whole vial as a starter



OK, thanks for the clarification Andrew. I could see that's what everybody was talking about. So I guess, just take my post as a reminder of what the conventional method is  I can see the slightly easier increased ease of starting the starter this way, but that is going to be offset by the increased number of slants you have to make. 

I think this is a worthwhile method if you brew fairly often with only a couple of types of yeast.

Steve


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## Sprungmonkey (7/5/08)

Steve Lacey - The only reason I make multiple slants is to give to a mate. We alternate buying yeast and I do the slants for him.


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## kram (7/5/08)

Sprungmonkey said:


> Steve Lacey - The only reason I make multiple slants is to give to a mate. We alternate buying yeast and I do the slants for him.


Don't buy WLP001, 830 or 833. I'll be slanting some up in the coming weeks.


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## ausdb (7/5/08)

Leigh said:


> From my lab days, the size of the magnet in the magnetic stirrer, the size of the stir bar, the speed of the stirrer and the viscosity of the solution all affect the stability of the stir bar...if you are not getting the stir bar to centre, try a smaller stir bar and/or start stirring slow and gradually increae the speed...even then, sometimes you can not get up to the maximum speed of the stirrer.
> 
> Flat bottom glassware assisted the stability...round bottom flasks needed quite small stir bars to stay stirring centrally.


Hi Leigh sorry I didn't see your reply till now. My stirrer is a home made jobbie out of an old 51/4" floppy disc drive with a belt driven platter, it only turns quite slowly about 360rpm put is not able start up from 0 rpm. I think the stir bar I am using is about 40mm long I will try a smaller one and see if it works any better
Cheers


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## quadbox (8/5/08)

I'd be quite happy to trade some WLP-023 (burton ale, aka wyeast 1275 thames valley) for good lager strains, or good belgian strains  This thread could so degenerate into a swap meet 

Steve lacey: Personally, I was only planning on having quite a few slants of burton ale and california ale, just because that's what I use 90% of the time. Will probably have ten each of those, and the remaining 30 vials I have will be used for say three each of more obscure or less used strains (a pilsner strain, a boch strain, a california common strain, a witbier strain, a trappist strain or two, coopers, an irish and a scottish strain ideally eventually  Oh my that hungry beast ambition )


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## Sprungmonkey (8/5/08)

Kram - I have San Fransisco Lager, Mexican Lager in slants , and Edinburgh Ale and Californian Ale still in White lab tubes ready to use.

Am using my first mexican lager slant in a "corona" style beer at the moment. See how it turns out.


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## mikelinz (8/5/08)

kram said:


> Don't buy WLP001, 830 or 833. I'll be slanting some up in the coming weeks.



I will have wyeast 2206 Barvarian in the fridge waiting for next time I brew, will slant it so will have available for a swap.


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## mikelinz (8/5/08)

~MikE said:


> should we perhaps set up a AHB wiki page where people post up their stocks of yeats. then we could just PM trade via mail etc
> 
> but yeah, i'm definitly up for it.



Can we just create a thread called "Yeast slant Library" (or something) probably in The Brew Shed > The Common Ground > Important Topics 

Not sure how to get it into "important topics" but the yeast farming thread is here. 

anyone know how we do it. My idea is we post what we have available and ppl PM to arrange a swap. Not sure how you keep the list up to date add and remove availability etc


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## ~MikE (8/5/08)

mikelinz said:


> Not sure how you keep the list up to date add and remove availability etc



that's why i was thinking a wiki page, it'd basically be AHB members listing their yeast stocks - only possible issue is working out an easy format for what would essentially be a giant list. each person would then be responsible for updating their section themselves.

the other option could be something similar to the recipe database...


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## newguy (8/5/08)

~MikE said:


> that's why i was thinking a wiki page, it'd basically be AHB members listing their yeast stocks - only possible issue is working out an easy format for what would essentially be a giant list. each person would then be responsible for updating their section themselves.
> 
> the other option could be something similar to the recipe database...



Not sure how hard this would be to create/code, but....

- Each user has their own inventory page, maybe sorted by dry/white labs/wyeast/other. Each known commercial strain has its own tick box. Users just tick what they have.
- Each user should also have a location or brew club tick box. Maybe sort by state so that it's not too "fine" of a location sort?
- The public page is sorted similarly, with dry/white labs/wyeast/other tabs/fields. Clicking on one would bring up an inventory of what is available, perhaps sorted by state/club, if sort by location/club is selected. Clicking on a particular available strain brings up a list of all users who have it in stock.

Sorry for sticking my nose in, but I sort through a lot of data and this breakdown method makes sense to me.


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## Moray (8/5/08)

I have just made my first slants and am wondering about my technique

I sterilised the tubes and caps in a microwave baby bottle steriliser, and quickly poured the boiling agar solution from my conical flask into the tubes and then capped them.

Is it really necessary to boil the agar solution once it's been added to the tubes, or should my dodgy method be ok?


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## ~MikE (8/5/08)

Moray said:


> I have just made my first slants and am wondering about my technique
> 
> I sterilised the tubes and caps in a microwave baby bottle steriliser, and quickly poured the boiling agar solution from my conical flask into the tubes and then capped them.
> 
> Is it really necessary to boil the agar solution once it's been added to the tubes, or should my dodgy method be ok?



so you microwaved your media (i'm assuming to boil agar and pasteurize) and chemical sterilized the tubes. that should be ok. if stuff grows, you'll need to go to a longer boil for your media - possibly even use a pressure cooker (like many people here do).


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## Zwickel (8/5/08)

~MikE said:


> so you microwaved your media (i'm assuming to boil agar and pasteurize) and chemical sterilized the tubes. that should be ok. if stuff grows, you'll need to go to a longer boil for your media - possibly even use a pressure cooker (like many people here do).


yeah, think so too, should be okay, but Id prefer a pressure cooker.

:icon_cheers:


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## Sprungmonkey (9/5/08)

shouldnt be any problems. Similar to the method I use. I Autoclave about 1L wort (with agar-agar) and then pour into sterile centrifuge tubes then cap. 121C doesn't let bugger all survive.\

Just leave the tubes at room temp for 4-5 days and watch to see if you get any growth. If you do then toss em before streaking your yeast.


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## edoeven (14/5/08)

hey guys, awesome thread  I plan to make a bunch of slants soon (i'll put up some pics) and start building up a yeast library - finally put some of those uni microbiology skills to use.... I wont have anything good to swap, but would anyone in melb have some interesting liquid strains they want to exchange for a few bottles of beer or something like that ?


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## tynian (14/5/08)

I slanted some 3068 the other day and it looks to have worked a charm.

Thanks so much for all the great info in this thread.


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## Linz (14/5/08)

Dont know if ive got the jist of this or not.....BUT.......

Can you make up say 30-50 'slants', then store them before innoculating them with yeast ??

Its just that if you're only going to need 4-6 slants of each yeast variety, 1 lt of wort/agar agar mix will fill more than the 4-6 you'll innoculate??


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## kabooby (14/5/08)

Yes mate you can. Thats what I have done

I have got about 20 blank slants waiting to be inoculated. When I decide to make a starter from some yeast that I currently have in bottles I will use some to inoculate the slants and go from there.

Dont order any vials. If you need some I ordered a 100 and I dont think I will use that many 

Kabooby :icon_cheers:


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## hockadays (14/5/08)

Kabooby,

I picked up some of the same vials that you have and made up 20slants today with 4g of agar to about 300ml of wort. Layed them down and it seems to have set quite well but I've noticed that if I rotate the vial the agar seems to slide around inside. It keeps it's shape and doesn't come loose but there is some movement, is this normal? They have also only been lying down for a few hours so i'll let them sit for 24hrs before I inoculate them.

hockers


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## Sprungmonkey (14/5/08)

shouldn't matter at all. some of mine do that after i streak them due to co2 production.


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## kabooby (14/5/08)

hockadays said:


> Kabooby,
> 
> I picked up some of the same vials that you have and made up 20slants today with 4g of agar to about 300ml of wort. Layed them down and it seems to have set quite well but I've noticed that if I rotate the vial the agar seems to slide around inside. It keeps it's shape and doesn't come loose but there is some movement, is this normal? They have also only been lying down for a few hours so i'll let them sit for 24hrs before I inoculate them.
> 
> hockers



I dont think it will matter. As Sprungmonkey said this can happen when you inoculate anyway

Kabooby :icon_cheers:


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## hockadays (14/5/08)

cool sounds like I'm on the right track,,,, B)


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## Cortez The Killer (15/5/08)

If someone is in the mood to organise a bulk buy for vials for slants 

Livingstone have the following for sale (I'd emailed them previously to get dimensions etc) - couldn't see anything else on their site that would be appropriate

TP50SU 50ML PP SAMPLE FLAT BTM UNLBLDCONT SCREW CAP STERILE 700/CT $149.16 (inc GST) 
Link: http://www.livingstone.com.au/catalogue.as...amp;cat_id=5292
Dimensions: 102mm high and 27mm diameter, polyethylene cap and polypropylene body.

The polypropylene tubes are autoclavable 
The polyethylene caps are autoclavable for 20 mins @ 121*C

Livingstone equates to 21.4c per tube
Proscitech equates to 37.4c per tube 
That's a massive saving of 16c per tube!

The only problem is that you need to buy 700 of the buggers

Links:
http://proscitech.com.au
http://livingstone.com.au

Note: I'm not putting my hand up to organise

No affiliation

Cheers


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## Interloper (15/5/08)

Hi all, I posted about my very amatuer yeast harvesting here on the forums so forgive the cross posting, but I'd like some expert advice.

I pulled 2 litres of my trub/slurry off into bottles on Tuesday night and have it stored in plastic in the fridge. It has separated into beer water on top and solid mud on the bottom - a good few centimetres of banded sediment.

Is this re-useable? Should I have been pouring off the beer water from the top? I opened it to let some carbon dioxide out this morning and it was pretty fizzy.

Is this worth trying to re-use? 

Any advice for cleaning this up or should I chuck it? (It was a Nottingham Danstar yeast)

Cheers


----------



## therook (15/5/08)

Interloper said:


> Hi all, I posted about my very amatuer yeast harvesting here on the forums so forgive the cross posting, but I'd like some expert advice.
> 
> I pulled 2 litres of my trub/slurry off into bottles on Tuesday night and have it stored in plastic in the fridge. It has separated into beer water on top and solid mud on the bottom - a good few centimetres of banded sediment.
> 
> ...



Have a look at this link

Link

Rook


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## mikelinz (15/5/08)

I got 50x30ml containers with lids for $31 including postage, you can get 100x10ml for about the same price.

link is here http://www.proscitech.com.au/cataloguex/on...p?page=l9#lch04

I got the PP with PP lids (LP9025-UU Vee bottom containers, PP, with PP screw cap, 30mL, unlabelled AUD17.00 /pk/50) as these are Autoclavable, so no need to leave the lid loose while they cool after boiling, i feel that this increases the chance of them getting infected and should work when I get a pressure cooker.

There is a description of the differant plastics and their properties/tolerances here http://www.proscitech.com.au/cataloguex/plastics.asp


They do drop their price to 15.30 if we order 10X50 but I suspect that the extra postage would negate the advantage of a bulk buy.

I suspect that the 10ml ones would do but went for the bigger ones as I felt they would be easier to work with. Anyone else got an opinion on the best slant size???

rgds mike


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## mikelinz (15/5/08)

ADDIT to my last the postage for 10x50 30ml jars is only 16.50 so probably there would be a saving of nearly $10 pp if we did a bulk buy. Any interest????

I would take another 50 and would be happy to coordinate. There is only a $6 saving (postage only) if the order is doubled.

rgds mike


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## hockadays (20/5/08)

Just thought I'd share my first go at slants and completion of yeast ranch. Very easy to do and better way then the previous I was using which resulted in mostly contaminated samples.


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## Hogan (31/5/08)

Greetings yeast culture's. 

I put together my first batch of slants today. They are currently solidifying and waiting for the inoculation stage. I have three questions:

1. After sterilisation the tubes, filling with the feeder solution and sealing - there is condensation in the tubes. Will this h2o have an adverse effect after inoculation? 

2. If so, is there a way to remove it?

3. When inoculating the slant should the growth medium be scratched to allow better grounding for the cells or is it only necessary to smear across the surface.


Cheers, Hoges.


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## Zwickel (31/5/08)

Hogan said:


> I have three questions:
> 
> 1. After sterilisation the tubes, filling with the feeder solution and sealing - there is condensation in the tubes. Will this h2o have an adverse effect after inoculation?
> 
> ...



howdy Hogan,

to 1.) thats just normal, after a couple of days the condens water will disappear, anyway it should be sterile and doesnt really matters.

to 2.) earlier in the thread Ive described a possible method to avoid an overly moisture in the vessels

to 3.) no, dont need to scratch, just smear across the surface or, like I do, roll a cotton wool swap over the surface.

Cheers :icon_cheers:


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## Hogan (1/6/08)

Zwickel said:


> howdy Hogan,
> 
> to 1.) thats just normal, after a couple of days the condens water will disappear, anyway it should be sterile and doesnt really matters.
> 
> ...




Thanks Zwickel. 

Cheers, Hoges.


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## Back Yard Brewer (1/6/08)

Picked up a couple of Agar plates from work on Friday. After reading this post and the fact that I opened up a brand new pack of 1084, I had to give it a bash. The dark patches are where I broke through the agar and hopefully not mould. The plates were left at room temp for 24hrs and have since been put into the fridge.

Back Yard Brewer


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## onescooter (1/6/08)

Hey, I really appreciate all the good info in this thread as I have been trying to move back towards liquid yeasts and this looks like a great way for me to do it. Couple of questions though.
1) If using the cotton swab method, do you sanitize it in any way.
2) Where do you get ethyl alcohol from as it seems like a nice way of cleaning.

If any one is looking into a bulk buy of test tubes I would be interested. May just order them direct but if there is bulk at cheaper prices I am keen.
Cheers.
Scott.


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## kabooby (1/6/08)

Hey Scott,

You can sanitize it buy steam. You can get ethyl alcohol from a chemist. They may sell you a bottle or you can get it in the way of mediswabs.

I haven't used the alcohol on cotton tips but have used it for a loop

Kabooby


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## Zwickel (1/6/08)

the professional way is to use a platinum loop, sterilized over a Bunsen burner (gas flame). Im using steam for the cotton wool. Dont use any chemicals or alcohol for that, it will kill the yeast as well.

Back Yard Brewer, Petri dishes are not really practical, since they are not hermetically closed. Petri dishes are more for a short term use.

Cheers :icon_cheers:


----------



## Back Yard Brewer (1/6/08)

Zwickel said:


> Back Yard Brewer, Petri dishes are not really practical, since they are not hermetically closed. Petri dishes are more for a short term use.
> 
> Cheers :icon_cheers:




Thought I would give it a trial for short term. Spur of the moment thing when I saw them kicking around work and just after I read this post. I want to try and use one in a couple weeks.

BYB


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## ~MikE (1/6/08)

onescooter said:


> 2) Where do you get ethyl alcohol from as it seems like a nice way of cleaning.



methylated spirits is 96-99% ethanol. is perfect for sterilization, just dilute to 70% for hands, surfaces etc, and leave as is for sterilizing a loop (as you want it to vaporize before looping cells)


----------



## TidalPete (1/6/08)

I used to wave my ss inoculation loop over a naked flame (cigaratte lighter or similar).
Don't bother with agar etc any more. I just either split a pack into 6-8 tubes or farm a used yeast into a similar number of tubes.
Good information re the metho. :icon_cheers: 

TP :beer:


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## Sammus (1/6/08)

Only just found this thread again 



Zwickel said:


> have you ever done it? you can see the difference very well. Put it in hot wort and youll get clumps, hard to dissolve. No problem in cold wort.


I have, and the agar just sunk as a powder to the bottom and never dissolved... Or do you mean to add it to cold water so it doesnt clump, then boil, during which it will dissolve?

I originally wanted to use this as a vegetarian gelatin substitute, and for this reason its not always possible.



> the stir bars are made to get autoclaved. Otherwise you could use a stirbar only once.
> After use Im sterilizing the bars in a steam pressure cooker.
> We have stir bars in our laboratory they get used since more than 10 years and steady autoclaved, they havent lost theire magnetism, not at all.
> 
> ...



Thats interesting, the magnets I play with say that if they are heated they loose their magnetism.. maybe they have to get hotter than an autoclave before that happens?


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## newguy (2/6/08)

All magnetic materials will lose their magnetism (it will actually be reset to 0) if heated above the Curie Temperature. This link lists the Curie Temperature of a few ferromagnetic materials down the right side of the page. To convert K to C, subtract 273 from the temperature in K. The warning for your stir bars may be because they're made out of a material with a low Curie point, or because the manufacturer is just trying to cover their ass. It probably has more to do with ass covering than anything else.


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## Zwickel (2/6/08)

thats right Newguy, but the materials gets back again its magnetism as soon as the temp drops below the Curie temp.

Cheers :icon_cheers:


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## newguy (2/6/08)

Zwickel, no they don't. What happens is that the magnetic domains within the material randomize at the Curie point. Unless the material is placed in a strong magnetic field as it cools, it's no more magnetic than an ordinary piece of iron. The randomization of the domains means that the net magnetization for the entire object is nearly zero because for every domain that points in one direction, chances are there is another somewhere that points in the opposite direction. The best you could hope for is that the bar would gain the magnetization of the earth's field, which isn't particularly strong, and chances are it would be at some odd angle to the bar instead of N at one end and S at the other.

The bar would still couple to a magnetic stir plate, but the coupling won't be as strong as for the original, unaltered, stir bar's field. This would translate into the bar losing coupling with the plate at a lower RPM than would otherwise be the case.


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## Back Yard Brewer (2/6/08)

newguy said:


> Zwickel, no they don't. What happens is that the magnetic domains within the material randomize at the Curie point. Unless the material is placed in a strong magnetic field as it cools, it's no more magnetic than an ordinary piece of iron. The randomization of the domains means that the net magnetization for the entire object is nearly zero because for every domain that points in one direction, chances are there is another somewhere that points in the opposite direction. The best you could hope for is that the bar would gain the magnetization of the earth's field, which isn't particularly strong, and chances are it would be at some odd angle to the bar instead of N at one end and S at the other.
> 
> The bar would still couple to a magnetic stir plate, but the coupling won't be as strong as for the original, unaltered, stir bar's field. This would translate into the bar losing coupling with the plate at a lower RPM than would otherwise be the case.




Way over my head h34r: :unsure: I think I will settle for a sterile spoon....

BYB


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## Sprungmonkey (2/6/08)

I use a inoculation loop and just heat over the stove (or with some metho and a candle). Get the loop red hot and let it cool for about 5 secs before streaking.


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## Hogan (2/6/08)

Zwickel said:


> Im using steam for the cotton wool. Dont use any chemicals or alcohol for that, it will kill the yeast as well.
> 
> 
> Cheers :icon_cheers:




Zwickel - After passing the cotton bud through the steam how long does it take the bud to cool down to a temp that won't kill the yeast cells when you dunk it in the inoculating yeast and then into the slant.

Cheers, Hoges.


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## Hogan (2/6/08)

Re the above post. 

I did some test runs using the kettle for steam and running the cotton bud across the spout for 15 seconds. Considering that steam temp would be in excess of 100c when it hits the bud, the tip of the bud cooled quite quickly. 

I am also thinking that as I am using a flame source to clean the air in the vicinity of where the transfer from primary yeast source to slant takes place, then, if the transfer tube is sterile and the receiving slant is sterile and the cotton bud is sterile, then all slants could be inoculated without having to re-steam the cotton bud after every transfer.

Any thoughts please.

Cheers, Hoges.


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## Franko (2/6/08)

Hogan said:


> Re the above post.
> 
> I did some test runs using the kettle for steam and running the cotton bud across the spout for 15 seconds. Considering that steam temp would be in excess of 100c when it hits the bud, the tip of the bud cooled quite quickly.
> 
> ...



Thats right hoges you dont need to re steam the bud after the transfer keep innoculating 

Franko


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## Zwickel (2/6/08)

Franko said:


> Thats right hoges you dont need to re steam the bud after the transfer keep innoculating
> 
> Franko


yeah, thats right.

Im using only one cotton tip for inocculating all slants.
When I have steamed one, I dont wait for cooling down, dip it straight into the yeast cake, there is so much yeast and the cotton tip will chill immediately, no worries.

:icon_cheers:


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## hoohaaman (1/7/08)

Steve Lacey said:


> 1) Why are all you people wasting a whole slant each time you brew? Just use a sterilized (flame or alcohol) loop or needle or whatever to pull out a blob of yeast from the slant and step up a starter going: 20 mL ... 200 mL .... 2000 mL (or less). And your slant will be good to use again tomorrow, next week or next year (conditional to the answer in part 2). The photos with rows and rows of slants seems unnecessary unless the user is about to culture up a dozen or so different strains. You should never need more than three slants of any variety, and even then two of them are reserves.



I have previously used the whole slant.After sterilizing with flame,I have no space in my slants to cool on agar.Could I cool in a Iophodor solution?Or would sterilizing with metho be a better/easier solution?

Cheers


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## Sprungmonkey (1/7/08)

hoohaaman said:


> I have previously used the whole slant.After sterilizing with flame,I have no space in my slants to cool on agar.Could I cool in a Iophodor solution?Or would sterilizing with metho be a better/easier solution?
> 
> Cheers



Just cool in the air - within 30cm of the flame. Should only take a few seconds. I actually dip in metho and lit off the flame wait 3 secs and innoculate from tube to slant. Therefore all surfaces of the loop are sterile.


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## hoohaaman (3/7/08)

Sprungmonkey,I don't mean for inoculating a blank slant.Rather using a sample from an established slant to prime a starter,instead of using the whole slant as a starter.

As Steve Lacey said,take some yeast from the slant to start,the starter

Cheers


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## Zwickel (3/7/08)

each time you open a slant, youll get some germs from the air into the vessel. 
Its easier and anyway safer, to use a slant only once.
Also it is much easier just to put some milliliter wort into the slant and leave it closed until one can notice some yeast activities, than to fiddle some yeast cells out of the agar and transfer into another vessel.

just my 2 cent.


Cheers :icon_cheers:


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## hoohaaman (3/7/08)

Thanks Zwickel,so better to make the number of slants you will use in a given period.Then to re-use a single slant more then once without Lab equipment?

I have enough space now,after changing to slants,but thought I may get away with using even less.

Cheers


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## Zwickel (3/7/08)

hoohaaman said:


> Thanks Zwickel,so better to make the number of slants you will use in a given period.Then to re-use a single slant more then once without Lab equipment?


yeah, thats the way Im doing it. The many slants Im always producing is not only to get used by myself, its also intended to get swapped with other brewing mates around here.
So Im well known around here, always to have some yeast in stock, its my hobby in the hobby :icon_chickcheers:


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## hoohaaman (3/7/08)

Haha cool,I also consider it a hobby inside the hobby


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## Sprungmonkey (4/7/08)

hoohaaman,
You can do it any way you want really. As Zwickel you increase the chance of contaminating a slant and using in a ruining a batch. I don't use slants more than once but there is no issue doing it if you use aseptic technique.
When making a starter as Zwickel said you can add some sterile wort to the agar slant and step up from there. I either do that or i metho and flame my loop and take a nice loopful of yeast and inoculate it into 10-20ml of wort, but you can do it anyway you want as long as it is aseptic. I find loop easier to use than cotton buds but that just me.


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## hoohaaman (4/7/08)

Thanks guys,I'm leaning more towards re-using a slant.But,then again the vials are small and cheap,so inoculating a dozen at a time doesn't phase me either.

I have a rather large yeast bank,changing to slants has freed up a large slice of the fridge.So I may as well be on the safe side and single use each slant.

I think I'd cry if I infected a batch due too poor yeast handling


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## hockadays (6/7/08)

After you use the slant can't you just wash out the agar of the tube clean it then use it again next time you make up a batch of blank slants?


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## Zwickel (6/7/08)

hockadays said:


> After you use the slant can't you just wash out the agar of the tube clean it then use it again next time you make up a batch of blank slants?


yes, of course you can, but.....
a brandnew and sterile tube cost only a few cent, depends on how many youre going to buy, so to clean and resterilize the tube is not worth the effort.

:icon_cheers:


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## Chad (19/7/08)

Has anyone given the LP9716 Tubes a go. I can see that I would be able to throw the tubes into a pressure cooker, but how will the caps go?
I see from this table that the caps would have to be a High Density PE, but the product description doesn't differentiate between LD and HD.

Has any found 10mm tubes to be too small anyway? Would the 30ml be a better option? I'm just trying to save space in my already limited fridge.


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## Vlad the Pale Aler (19/7/08)

Chad, I bought 100 of the LP9716 tubes. Too small and fiddly, go with the 30ml. I boiled the tubes and caps together with no problems.

I've had a go at slanting and can't decide if its really worth the stuffing around, with the increased risk of infection and failure.
I will use the tubes to split up packs of fresh yeast and reanimate from them, much easier and safer.


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## mikelinz (19/7/08)

Chad said:


> Has anyone given the LP9716 Tubes a go. I can see that I would be able to throw the tubes into a pressure cooker, but how will the caps go?
> I see from this table that the caps would have to be a High Density PE, but the product description doesn't differentiate between LD and HD.
> 
> Has any found 10mm tubes to be too small anyway? Would the 30ml be a better option? I'm just trying to save space in my already limited fridge.



I went with LP9025 for 2 reasons 30ml, easier to work with (yup uses a bit more medium but costs bugger all) and both lid (PP) and container (PP) can be autoclaved, for when i get around to getting a pressure cooker. You can boil/pressure cook these with lid screwed on and they won't collapse when they cool down. I offered to do a bulk buy but didnt get any feed back. Still could.


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## hockadays (19/7/08)

mikelinz said:


> I went with LP9025 for 2 reasons 30ml, easier to work with (yup uses a bit more medium but costs bugger all) and both lid (PP) and container (PP) can be autoclaved, for when i get around to getting a pressure cooker. You can boil/pressure cook these with lid screwed on and they won't collapse when they cool down. I offered to do a bulk buy but didnt get any feed back. Still could.




Same here and the 30ml are easy to step up in when you want to use them.


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## Chad (19/7/08)

Thanks guys, 30ml it is.


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## mikelinz (25/7/08)

Vlad the Pale Aler said:


> Chad, I bought 100 of the LP9716 tubes. Too small and fiddly, go with the 30ml. I boiled the tubes and caps together with no problems.
> 
> I've had a go at slanting and can't decide if its really worth the stuffing around, with the increased risk of infection and failure.
> I will use the tubes to split up packs of fresh yeast and reanimate from them, much easier and safer.



I guess you are missing the one point of stanting. If the slant is contaminated you can see it as all non yeast populations will grow visually differant. There is no visual check if you simply split you yeast while there is potentially an increased risk of contamination. Also slanting allows you to grow on populations whereas just spliting the pak means that you will still have to buy new ones just not as often.

rgds mike


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## Vlad the Pale Aler (25/7/08)

mikelinz said:


> I guess you are missing the one point of stanting. If the slant is contaminated you can see it as all non yeast populations will grow visually differant. There is no visual check if you simply split you yeast while there is potentially an increased risk of contamination. Also slanting allows you to grow on populations whereas just spliting the pak means that you will still have to buy new ones just not as often.
> 
> rgds mike




Thanks Mike, but at $9.80 for a smackpack, as opposed to the time it takes to play with slants, I'll take the risk.


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## schooey (25/7/08)

I read a lot about this topic now, but I still have a couple of questions. Maybe they have been covered off, but I can't find where, so anyway...

I read somewhere that a a few people make up their media by boiling some LME, Agar Agar and Yeast nutrient and then put it in their vials that have been pressure cooked. From the impression I got, they then pressure cook the whole lot again to sterilise? If this is the case, do you do that with the lid on or off the vial? and if it is off, how do you stop all the condensate in the pressure cooker filling up in the vials?

:blink: Maybe I'm way off here...


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## braufrau (25/7/08)

OK .. haven't done it yet but ...

put the unsterilised vials with a agar/malt solution and pressure cook.
The lid has to be on or its not pressure cooking, and it will only get to 100C.


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## schooey (25/7/08)

:lol:.. I didn't mean does the lid need to be on the pressure cooker, I get that bit, I meant does the lid need to be on the vials _in_ the pressure cooker to avoid them fillng up with moisture?


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## MCT (25/7/08)

schooey said:


> I read a lot about this topic now, but I still have a couple of questions. Maybe they have been covered off, but I can't find where, so anyway...
> 
> I read somewhere that a a few people make up their media by boiling some LME, Agar Agar and Yeast nutrient and then put it in their vials that have been pressure cooked. From the impression I got, they then pressure cook the whole lot again to sterilise? If this is the case, do you do that with the lid on or off the vial? and if it is off, how do you stop all the condensate in the pressure cooker filling up in the vials?
> 
> :blink: Maybe I'm way off here...




Good question Schooey, I've been wondering this too.
All the guides I've read have said to sterilize with the lids off the vials, which has lead to alot of condensation forming drops at the bottom of the slanted media.
My slants have ended up looking like this:


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## braufrau (25/7/08)

LOL!

I think I'm suffering from sugar poisoning! Sorry about that.

OK .. I don't know .. but I'm planning on just sitting them on top. 
Then I was going to screw them on with sanitised hands.

This ppt thingy about autoclaving says to vent lids on solutions. linky
and this link on propogating orchids also says to leave them on loosely.
linky

Better wait 'til someone who knows answers.


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## Sprungmonkey (27/7/08)

MCT - why don't you just tip the condensate out before innoculating the slants?


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## braufrau (27/7/08)

Well I've done mine and I put the lids on top, but didn't screw them on until they were done. Aren't they pretty?






The bit of fencing wire was autoclaved too as well as the little jar for putting aside some starter for inoculation in.


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## Vlad the Pale Aler (27/7/08)

braufrau said:


> Well I've done mine and I put the lids on top, but didn't screw them on until they were done. Aren't they pretty?
> 
> 
> 
> ...



During my short slanting career I used a large darning needle, looped end, to inoculate the slants, worked quite well.


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## braufrau (27/7/08)

Vlad the Pale Aler said:


> During my short slanting career I used a large darning needle, looped end, to inoculate the slants, worked quite well.




That would certainly be more elegant! 
I looked through my sewing box but couldn't find one quite the right size then hit upon the fencing wire idea. 
I think that's what's known as "a bit agrigultural!".


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## mickoz (27/7/08)

I've only ever used an electric steamer to sterilise my slants after filling them with malt jelly. Seems to work fine as I have had no infections and it is a whole lot easier than mucking with a pressure cooker.

I will take some photos next weekend and post them here as I have run out of clean slants and need to make more. I actually use specimen jars, they're a bit easier to handle I think.

Mick


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## Chad (31/7/08)

I'm now another step closer to preparing my slants  .
Found a very decent priced pressure cooker from Victoria's Basement which I just ordered. Went for the slightly better model as I want to use it for cooking as well. I don't know what this company is like as this my first purchase from them.


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## hoohaaman (31/7/08)

mikelinz said:


> I guess you are missing the one point of stanting. If the slant is contaminated you can see it as all non yeast populations will grow visually differant. There is no visual check if you simply split you yeast while there is potentially an increased risk of contamination. Also slanting allows you to grow on populations whereas just spliting the pak means that you will still have to buy new ones just not as often.
> 
> rgds mike



Well mikelinz,I didn't want 100 plus slants in the fridge.So I took your advice and culture off one.Took all precautions,flame above working area ect.

First reculture from slant success,contaminate original slant seems not.I think the single slant(have 3 backups)will last many brews.Will of course do visual,smell and taste to proof.But I'm a convert and will be buying proper equiupment.

Thanks


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## mikelinz (1/8/08)

Vlad the Pale Aler said:


> Thanks Mike, but at $9.80 for a smackpack, as opposed to the time it takes to play with slants, I'll take the risk.




Where are you buying them for $9.80 Vlad


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## newguy (1/8/08)

I believe that Craftbrewer is selling them for $9.80.

Edit: just checked; yes, it's Craftbrewer.


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## Benniee (8/8/08)

Thought I would stick up a pic of my first slant. It's done in a baby food jar as I didn't have any tubes at the time.

I inoculated it with some US-05 yeast from a harvested slurry - I know it's a dry yeast but I wanted to test my method was sound.

To my untrained eye it seems ok. It took a while to get going as the weather has been pretty cold, and the condensation has pooled a little bit in the bottom of the jar.

The next step will be to make a starter from it, and taste test the result.

Benniee


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## Sprungmonkey (8/8/08)

absolutely nothing wrong with doing it this way Benniee - just takes up a bit more room than a tube. looks good.


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## Chad (9/8/08)

I tried a first run at some slants today, and based on my results I have a few questions.

1. Are you pressure cooking the wort + vial together?
2. If so, are you screwing the lid on tight? I didn't (the lid was screwed on and then back 1/4 turn) and found that the wort was bubbling back out.
3. If the lid is on tight, will the container deform, or is there an equal pressure between inside of vial and outside of vial to negate this?


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## Zwickel (9/8/08)

Sprungmonkey said:


> absolutely nothing wrong with doing it this way Benniee - just takes up a bit more room than a tube. looks good.


+1 subscribe
yeah, when I was starting with yeast farming, I did it exclusively this way. 
The advantage is, one can start with a bigger amount of wort, means your starter gets ready to pitch earlier.

Disadvantage is only it takes more space.

Keep on doing....Cheers :icon_cheers:


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## braufrau (9/8/08)

Chad said:


> I tried a first run at some slants today, and based on my results I have a few questions.
> 
> 1. Are you pressure cooking the wort + vial together?


yep.



> 2. If so, are you screwing the lid on tight? I didn't (the lid was screwed on and then back 1/4 turn) and found that the wort was bubbling back out.


Nope. Just sitting it on the top. I only 1/4 filled the tube.



> 3. If the lid is on tight, will the container deform, or is there an equal pressure between inside of vial and outside of vial to negate this?


There is some risk .. I'm not sure how much .. that the container may implode. That is why safety instructions for an autoclave say not to have the lids one. But I think an autoclave gets higher in P &T than a pressure cooker.


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## Sammus (9/8/08)

I do mine as like canning. Pressure cook the vials with the lid on, but until it only just starts to tighten. It's loose enough to let the pressure out when its hot, but as it cools it vacuums itself shut. I get shocking condensation but *shrugs*.

Only problem I'm having is plating to isolate cultures...I've been having bad trouble with infection - the same prep as I use with slants, but my petri dishes get covered in mould, and my slants stay clean *shrugs*...


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## Benniee (9/8/08)

Sammus said:


> Only problem I'm having is plating to isolate cultures...I've been having bad trouble with infection - the same prep as I use with slants, but my petri dishes get covered in mould, and my slants stay clean *shrugs*...



I haven't used petri dishes yet, but some stuff I've read mentions that they're not airtight, and usually need a bit of tape to seal the two parts together and keep air out. Are the dishes growing mold after you inoculate them or just while they're sitting there as blanks?

Benniee


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## ~MikE (10/8/08)

Sammus said:


> Only problem I'm having is plating to isolate cultures...I've been having bad trouble with infection - the same prep as I use with slants, but my petri dishes get covered in mould, and my slants stay clean *shrugs*...



first, you need to be anal when using petri dishes, they have a much larger surface area and opening than slants and will get infected easier. make sure you're getting the pre-sterilized dishes. once you've made them up, seal them. otherwise, even if you keep them in the fridge, fungi and bacteria will find their way onto them and will grow as soon as you go to use it. when using them, be very sterile. 70% ethanol your hands and bench. work near an open flame. flame your loop (preferably till it's glowing. avoid having the petri dish open any longer than you need to. seal it up again once innoculated.



Benniee said:


> Thought I would stick up a pic of my first slant. It's done in a baby food jar as I didn't have any tubes at the time.


that's a lot of media for slantage. if you're using a larger tube/jar, you don't really need to have it slanted. slanting the media is for increasing the media's surface area in a narrow tube. i sometimes use the 30ml specimen containers (similar to urine sample containers) - enough media to cover the bottom is plenty.

hope this helps.
-mike


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## Chad (10/8/08)

Chad said:


> 2. If so, are you screwing the lid on tight? I didn't (the lid was screwed on and then back 1/4 turn) and found that the wort was bubbling back out.


Just did a 2nd batch today with much better success and thought I would share. This time I put the lids on loose, and didn't have any troubles with the wort bubbling out.

My theory is that with my first batch, the lids weren't loose enough to let the gases escape fast enough, putting more pressure inside the vial, raising the temp. and making it boil violently. With the caps loose, the gas could escape and keeping everything inside the vial a bit more calm.


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## Benniee (10/8/08)

Chad said:


> Just did a 2nd batch today with much better success and thought I would share. This time I put the lids on loose, and didn't have any troubles with the wort bubbling out.
> 
> My theory is that with my first batch, the lids weren't loose enough to let the gases escape fast enough, putting more pressure inside the vial, raising the temp. and making it boil violently. With the caps loose, the gas could escape and keeping everything inside the vial a bit more calm.



So did you just open up the pressure cooker and tighten the caps once it had cooled a bit? I know that's a pretty obvious question but I'm wondering how cool you let the pressure cooker get.

Benniee


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## Benniee (10/8/08)

~MikE said:


> that's a lot of media for slantage. if you're using a larger tube/jar, you don't really need to have it slanted. slanting the media is for increasing the media's surface area in a narrow tube. i sometimes use the 30ml specimen containers (similar to urine sample containers) - enough media to cover the bottom is plenty.



Yeah, I realise that tilting the jar was probably unnecessary, but I wanted to to a true test run on making up a "slant".

I've got some 30mL tubes now so my next go will be in some smaller containers.

Benniee


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## Chad (10/8/08)

Benniee said:


> So did you just open up the pressure cooker and tighten the caps once it had cooled a bit? I know that's a pretty obvious question but I'm wondering how cool you let the pressure cooker get.


Once the pressure cooker was finished, the pressure was released immediately and opened. The caps on the vials were then done up tight.


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## mckenry (26/1/10)

Just thought I'd let all know about my latest reculture. I had a Wyeast 1084 Irish Ale. Cant even remember when I bought it, though it would be late 07 or early 08.
I took a vial from a brew in oct 08 and it has been sitting in my bar fridge since.
So, here's my method of reculturing for beginners, which is so simple.

Day 1 I took the sample out of the fridge to warm to room temp. Same time I boiled 50g of DLME in 500mL water for 10mins then put in a flask in a fridge (had a lid, but you could use glad wrap)

Day 2 Took the DLME mix out of the fridge and let it come to room temp. When room temp achieved, mixed the two and shake vigerously and insert bung and airlock. Swirl whenever I think of it.

Day 3 High krausen achieved and bubbling slowing. Made another starter - this time 100g in 1L water and same process followed.

Day 4 Poured off excess liquid, leaving yeast layer at the bottom. Smelt and tasted the liquid. No problem there. Mixed the latest DLME starter with the yeast and shaked vigerously etc.

Day 5 High krausen achieved and let to settle right out. Read about best to leave for 18 hours after fermentation, to build up food reserves.

Day 6 Pitched to 1.065 wort. 12 hours later the fermentation is in full swing at a temp controlled 19C.

Easy.


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## mckenry (15/10/11)

mckenry said:


> Just thought I'd let all know about my latest reculture. I had a Wyeast 1084 Irish Ale. Cant even remember when I bought it, though it would be late 07 or early 08.
> I took a vial from a brew in oct 08 and it has been sitting in my bar fridge since.
> So, here's my method of reculturing for beginners, which is so simple.
> 
> ...



Weird - I went looking for a thread like this to let other brewers know about my latest culture and find myself as the last poster, more than 18 months ago.
Turns out the yeast strain is the same one!
Which means this particular strain is 4 years old and only been used a handful of times.
I made a robust porter wort a while back, no chilled. The cubes have been in the garage for a while now. Decided it was time to brew them for next winter.
[These days my method is slightly different to the above. I now own a 5L flask & a stir plate and I get my starters as drainings from my MT]
I grabbed the vials from my bar fridge - dated Jan 2010. 
Boiled 4L of starter from my robust porter, directly in flask.
When temps met up, pitched the contents of the vials, and plonked on the stir plate for 24 hours.
No visible signs of krausen after another 24 hours, so just let nature do its thing.
The yeast layer at the bottom was growing every day, so I wasnt worried.
After 6 days I took the SG. Had gone from 1.040 to 1.008, so I pitched it after pouring off the liquid.
After one week in the PV the krausen ring has subsided, but evidence shows 150mm of krausen at some stage.
Glad wrap is still straining to burst, so its still fermenting.
Just goes to show that you can store for a long time and renewing the cultures, while it will keep the viable cell count up, isnt as necessary as some will have you believe.
Everything is going well, sample is good.

Note - all I ever do is grab some sludge from the bottom of the PV, into a few 50mL vials. 

mckenry


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