Guide to Making a Frozen Yeast Bank

Williams 728
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butterpants said:
You're joking right?
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I never asked, WHAT ebay was.
I said, WHERE.
Given the fact that I am in mainland China (shown next to my nickname) this isn't exactly a joke.
Ebay is a no go - it's no problem ordering there but shipment will cost stupid money and why bother if 90% of all sold on ebay is made somewhere a couple of blocks away.
So I shop at taobao.com, a (should be the) internet trade portal of China.
 
I only use the mobile app.... I can't see details like user location, avatars, sigs, etc without digging for them. My bad.

Yes, China does complicate things. Could be worse.... you could be in North Korea....
 
butterpants said:
Could be worse.... you could be in North Korea....
Click to expand...
Huhh, in my nearly 20 years in China there were times I loved her, liked her, did both, or did neither. But I'd never say it's as bad as to be remotely compared to NK. Hey, just from a brewer's point, what about a 13-gallon stainless pot (2.5 mm thick no crap monster) costing 23 bucks delivered to my door?
And take it from me, all this internet censorship and alike, when you think of 1.5 bln guys (talking people, not yeast cells!) that are enjoying some 30-years lasting peace period for the first time in like 500 years... could be worse, oh yes it could.
:)
 
POTDA
Just seeing this now... GQT gets my vote for post of three days ago.
 
Alright looking to add a frozen bank as redudency for long term storage of mother cultures in addition to the short term storage of liquid media via yeast starter harvesting.

What storage containers are everyone using? I do small batches, and am in an apartment, so probably looking at the 1.5-10 ml range. Needs to be a leak proof seal, autoclavable.
 
Get gamma irritated screw top pcr reaction tubes. Seals better than flip top and pre-steralized.
 
Just set up 5 frozen tubes of 510, and 570. Mixed up two nights ago, and am ready to freeze them.

Is the current mindset to freeze quickly, freeze slowly, or don't worry about it?
 
Don't know if it's too late but here use freezable Breast Milk Bags. My wife Breast feeds our babies. The bags are for storage and freezing of breast milk. They need to be steril from the factory. They have a double Ziploc and we've never had one leak. They are a bit big but you can store whatever amount you want. I squeeze out most of the air and freeze. No pressure cookers or sterilizing and I've never had a bad batch. Just use a new bag each time! A box of 100 is about $15-$20 but for me that box will last a long time.
 
I'm sure that works quite nicely for pitchable volumes. I prefer my frozen banks to be on the small size though so I can have a bunch of strains and not take up the entire freezer.
 
The bags are 6oz but I only put about 40ml at most in a pouch. I squeeze the air out so they dont take up space. What I do is freeze them flat and file them in a box / cooler like files because they're so thin. I have 5 yeast varieties in my freezer now and about 5-7 pouches each. That's about 30 pouches in a 8x6x6 cooler in my chest freezer. I write right on them what they are so no labels needed. I guess I could do pitchable volumes if I washed my yeast and used that, but I make starters and split that into smaller amounts. To each their own. I'm just telling what I do and works for me. If my idea works for someone else, great! If not, that's fine too.
 
Kwitty said:
The bags are 6oz but I only put about 40ml at most in a pouch. I squeeze the air out so they dont take up space. What I do is freeze them flat and file them in a box / cooler like files because they're so thin. I have 5 yeast varieties in my freezer now and about 5-7 pouches each. That's about 30 pouches in a 8x6x6 cooler in my chest freezer. I write right on them what they are so no labels needed. I guess I could do pitchable volumes if I washed my yeast and used that, but I make starters and split that into smaller amounts. To each their own. I'm just telling what I do and works for me. If my idea works for someone else, great! If not, that's fine too.
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This sounds great! My wife is breast feeding too, so I have a ton of these pouches. Your just freezing the yeast? Your not using any glycerine or anything to prevent cell damage to the yeast when freezing?
 
No! I'm still using the glycerin and doing all the same steps to freeze the yeast. I'm just not using reusable vials and sterilizing them each time. I am using the breast milk bags instead of the vials. You still need to prep the yeast, it's just a different container.
 
I dilute the glycerin with water to a 20% mixture and boil it in the microwave for 5 minutes. It may raise the percentage a bit but it's OK. I mix the glycerin mixture 50/50 with yeast slurry. That cuts the glycerin in half so we now have approximately 10% glycerin. Probably slightly higher with the water that evaporated in the boil.
 
Here is my go at frozen yeast banking it seems to have been a success. Check it out if your interested in seeing how its done.

[ame]https://www.youtube.com/watch?v=iDIwwnFFHLQ[/ame]
 
Jason I liked the video. Very off the cuff with decent layman's explanations of why you're doing what you're doing. My 2 cents....

1) Might look into adding a small amount of citric acid to the glycerine/DI water mixture. Helps with oxidation. Isotonic saline might be better than DI water too.

2) Keep an eye on those tube caps. The PP ones (not resin) can sometimes warp post autoclave. Wrap the tops in parafilm or electrical tape before freezing.

3) You go through all that trouble to sterilize then open all the tubes at once and expose them to the air? Don't do this... one at a time. Also light a candle/alcohol lamp and work in close proximity to it. The updraft will keep falling particulate (dust), which carry bacteria and wild yeast.

4) That Cinnamon Toast Crunch box in the background was calling my name the whole time....

Thanks for the vid
 
Hey,
I slowly reading this thread and a book " Yeast: The practical guide to beer fermentation" by Chris White and Jamil Zainasheff .
When I relocate my knifemaking workshop from Basement to garage, I have 2 clean rooms to use with steady 18*C in winter and summer.

In one I want to make very nice Yeast laboratory and bank.
In second home brewery.
Centrifuge, autoclave, laminar hood, -80*C freezer, 2*C - 45*C incubator with magnetic stirring plates ,microscope with mounted DSLR , fridge should be ok for that ? ( I know for home is overkill, but I like this type of precise work :D )

Should be a good lab for taking clean wild strains of yeast?
(And a make nice amount of micro-starters from yeast got from store :D )
 
Super overkill for a personal bank.

Keep it simple, plating for isolation cultures and ensuring purity + frozen bank.


No need to spend all that money and equipment if it's just for you and maybe some friends. There's no way you will ever overcome that investment with personal or even homebrew club usage.

If it's a pro setup, then sure MAYBE.
 
But I will get that stuff from my Aunt :D
She is microbiologist, and she want to give me her old stuff, but full functional :D (Centrifuge, Autoclave, small laminar hood and freezer)
I will only need to buy good microscope and build mentioned incubator ( from fridge ;) )

Hmm investment...
I already spend around 25.000$ on my knifemaking workshop ( but I'm a full-time knifemaker)
 
I like the cut of you jib fella.

.... but that level of sophistication wouldn't have stirplates in an incubator.... it would be shakers.

Orbital shakers are better than stirplates... less shear on cells. At the homebrew growth scale I don't think it matters much cuz it's pretty easy to pitch high but as James Bond mentioned, shaken is always better than stirred.
 
Is possible to step-up Lactobacillus into bigger amount (clean strains - brevis etc. ) and make a 1,5ml micro-starters for banking ?
From 1 pack then I can make nice amount of starters with clean culture ( L. Brevis etc. are pricey here >.>)

From what I reading, with time and practice I'll be able to isolate even bretts from wild forage ?
 
You can slant lacto. I use MRS agar. LAB grows like friken wildfire on this stuff.

I have no experience with freezing small amounts and reviving them. I'm sure someone here has.

Stepping up is easy and you need a suprisingly small amount of total growth media because bacteria are small and have a badass growth curve. Growing up in MRS media may not be optimal (well it WILL be for cell counts.... but it tastes n smells awful). You will want to propigate in a mixture of DME, Glucose, DAP and CaCO3.

http://sourbeerblog.com/lactobacillus-2-0-advanced-techniques-for-fast-souring-beer/

Dr Lambic has a big writeup on it. Super detailed
 
I'm pretty sure the American Sours book says you can just feed your lacto and pedio with apple juice every couple of weeks and it will keep. No need to freeze or slant/plate. You can treat them like a sour dough starter. Just keep it fed.
 
Breadontap said:
I'm pretty sure the American Sours book says you can just feed your lacto and pedio with apple juice every couple of weeks and it will keep. No need to freeze or slant/plate. You can treat them like a sour dough starter. Just keep it fed.
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Problem with that is growth and longevity can be self-limiting based upon how low the pH goes.
 
Your right. It did mention adding chalk to keep the ph in check with consistent decanting and feeding.
 
I'm a white labs guy myself but really who ever has the type of Brett or lacto you want. Look around the American sours book suggested http://www.eastcoastyeast.com/

I've never used them personally. It looks like they have a several blends.

I'm curious how often you plan to use lacto? I only use it when brewing a Berliner.
 
Goodbelly Caps - L. plantarum

White Labs - L. brevis

All u need.... Brett... like a million good options
 
FWIW white labs lacto is pretty well known to contain yeast, and so it's not a good option for sour worting or sour mashing. Combine that with their very low cell counts for lacto, and brett, and I don't recommend buying from them for those purposes.

If you can get it, omega lacto blend is amazing.
 
If you're proping it yourself, cell counts don't matter... growth is exponential and rapid.

The Omega lacto blend is plantarum and brevis if I remember correctly... hence why I mentioned the two. Blend them yourself!

WL yeast has lacto in it too... I'm glad someone else noticed this QC issue.
 
oh and if you propigate your LAB at a high enough temp the yeast don't survive (for the paranoid and people who can't isolate)
 
butterpants said:
oh and if you propigate your LAB at a high enough temp the yeast don't survive (for the paranoid and people who can't isolate)
Click to expand...

I read someone tried this on the MTF group like two weeks ago, can't remember if it was successful or not. IIRC it was incubated at ~120-140?
 
140 sounds pretty damm high. Not sure the LAB is gonna like it. Funk guys could have, I'm not aware of the experiment.

I think 120F is a nice place to start.
 

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