Guide to Making a Frozen Yeast Bank

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Awesome writeup! I have a few questions though....

I happened upon a great deal on a centrifuge ($50), but I'm
I'm having the hardest time finding glass 15ml centrifuge tubes. The only places that sell them seem
When I put the yeast in the freezer, do I use agar? I've read so many conflicting guides on this aspect that I'm very confused.
 
Agar is used when you're making slants or plates, which get stored in the refrigerator. Freezing is a different process, needing a cryoprotectant like glycerin/glycerol.

I got my vials at Cynmar.com. I'm sure they would have centrifuge tubes.
 
Agar is used when you're making slants or plates, which get stored in the refrigerator. Freezing is a different process, needing a cryoprotectant like glycerin/glycerol.

I got my vials at Cynmar.com. I'm sure they would have centrifuge tubes.

Thanks
I checked Cynmar, and the cheapest I could find were the 10ml ones.
Linky

Here's their selection

A great site is the lab section of sciplus.com. They have plastic 15ml centrifuge tubes for packs of10 for $3.50. I'm considering these, but I couldn't use them in a pressure cooker though. Maybe just star san the hell out of them and hang em upside-down?
 
Just a note for those who may want to save money. You can get away with buying plastic microfuge tubes (~1.5 mL) online. They're tiny, lab safe(-80*C), and they're plentiful. So you can just keep your glycerol stocks in those tubes and dispose of it after you make your starter. Then make a new one before you pitch you yeast.

Just a thought.
 
Slowly building my yeast bank.

Here's what I am doing; every time I am scheduling a brew day I pick a recipe with a yeast I really want to save, make a starter, put 8 to 10 viles into yeast bank and pitch the rest on the recipe wort.

So far I have
Wyeast 1056 California Ale 10 vile
Wyeast 1038 London Ale III 8 vile

Up next:
PacMan
Denny's Favorite
 
Slowly building my yeast bank.

Here's what I am doing; every time I am scheduling a brew day I pick a recipe with a yeast I really want to save, make a starter, put 8 to 10 viles into yeast bank and pitch the rest on the recipe wort.

So far I have
Wyeast 1056 California Ale 10 vile
Wyeast 1038 London Ale III 8 vile

Up next:
PacMan
Denny's Favorite

If they are vile, don't waste the vials on them. :cross: :D ;)

Sorry couldn't resist.
 
Thanks for the helpful writeup. My question is, is it necessary to do a small starter as an intermediate step to using one of the vials, or can I pitch the vial into a larger starter?

Time for me to build a stir plate.
 
With beer (starters and your carboy), you want to get the alcohol above 2% as soon as possible. This protects it from bacterial infection.

So, although you can pitch small into large, it might take several days to ferment out. During that time you are putting room-temp sugarwater at serious risk of bacterial growth. By starting small, you'll get that 2% pretty quickly, any bacteria present will (hopefully) be poisoned, and then you step up again.

I'm not sure, but I think if you pitch a vial into a 4L starter you'll get the same number of cells as if you had stepped it up.
 
Can somebody please expand upon this?

"If you don’t have a stir plate, make your starter in two rounds. For example, start with 3 cups of wort, ferment it out, and then add another 3 or 4 cups of wort to really build up the yeast."

How do I know when it is fermented out and do I just add the second batch of wort straight on top of the existing?
 
Can somebody please expand upon this?
Sure…
"… ferment it out, … "
This was a poor choice of words. Nobody wants to wait 1 to 2 weeks for a starter to be ready. I would wait till the yeast colony quits growing and gets to work. High Krausen is a good sign they are hard at work. Also be sure to add more nutrient and aerate the fresh wort so the yeast colony can begin growing again.

If you never see Krausen then take hydro readings. If the readings never drop, you have dead yeast.

IMHO, let the readings drop to around half what the OG was.

…and do I just add the second batch of wort straight on top of the existing?
Yes. Or pitch the existing starter into the new starter wort. Same thing.
 
Thanks Beer_Guy.

What size pressure cookers are you guys using? Is a 5qt one big enough? I am looking to pick up a cheap one online.
 
Thanks Beer_Guy.

What size pressure cookers are you guys using? Is a 5qt one big enough? I am looking to pick up a cheap one online.

If your just using it to sterilize your vials/glycerin/tools etc. it should be fine, although depending on length a pipette might not fit. However if you later want to pressure can jars of sterile wort to make starters with, a larger pressure canner will be more versatile. Keep in mind the size is based on the amount of liquid it holds, not the number of jars so if you want to can 6 -7 quart jars it takes about a 20-24 quart canner for them to fit. Try your local Goodwill thrift store or similar for a good price.

Here is a 23 quart http://www.amazon.com/dp/B0000BYCFU/?tag=skimlinks_replacement-20

holds 7 quart jars
 
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Do you guys sterilize your Glycerin? I've just been sterilizing the vials, the ssteel baster. soaking the caps in star san because sterilizing the caps shrunk the seal rendering them leaky
 
I spent a lot of time and money on this yeast bank project. I eventually stockpiled about 7 strains of yeast. Then I moved and all of it thawed and was warm for a couple of weeks. I never did even brew a batch from cultured yeast... :(

No regrets. I'll rebuild my library.
 
I think I'll start using some of mine now even though I only have 3 strains..I have 10 vials of each so no room left to really add much more in terms of # vials.
Have London Ale, Cal Ale 1056 and Pacman.just brewed a Dead Guy Clone with Pacman today from the starter so I'll have to think about what to brew next.

That's too bad about your yeast bank going bad..lots of work..
 
I spent a lot of time and money on this yeast bank project. I eventually stockpiled about 7 strains of yeast. Then I moved and all of it thawed and was warm for a couple of weeks. I never did even brew a batch from cultured yeast... :(

No regrets. I'll rebuild my library.

Ouch. At least you have all the hardware already, so only have to invest in the consumables.
 
I think I'll start using some of mine now even though I only have 3 strains..I have 10 vials of each so no room left to really add much more in terms of # vials.
Have London Ale, Cal Ale 1056 and Pacman.just brewed a Dead Guy Clone with Pacman today from the starter so I'll have to think about what to brew next.

That's too bad about your yeast bank going bad..lots of work..

So far I only have 5 vials of one strain :) Fat Tire ale yeast. Will probably add Pacman next even though the LHBS sells the Brewcraft Rogue licensed kit and the Pacman yeast all the time.
 
How big of a pressure cooker is everyone using for this? Do I need one with the gauge on it?

I started with a small 6 quart one without gauges and it got the job done, I only recently updated to a big fancy one when a friend moved and gave me his.
 
Is there any reason why the glycerine mixture cannot be put in the vials and then microwaved for sanitation?

microwaving isn't great for killing bacteria... the way it does it is with heat and you really need to maintain a high heat for several minutes, which idk if a microwave can do all that well.
 
microwaving isn't great for killing bacteria... the way it does it is with heat and you really need to maintain a high heat for several minutes, which idk if a microwave can do all that well.

Gotta call nonsense. If the water is boiling, it is 212F. What temp do you think you get in a pot?

Pressure cookers can get to higher temps. A pot cannot.
 
Gotta call nonsense. If the water is boiling, it is 212F. What temp do you think you get in a pot?

Pressure cookers can get to higher temps. A pot cannot.

Kinda confused... I don't really see how what you're saying is relevant to calling it nonsense.

I wasn't talking about pots...
 
Anyone else find a good source for equipment? Cynmar has a $25 minimun order but they do not sell pipettes or corning beakers. The local place in Denver sells the vials but the caps are not autoclavable.
 
I've frozen about 30 vials..but have not reconstituted any yet.The vials seems to be sealing properly.

I have some Denny's Favorite 50 on the way that I will be putting into the bank.
Once that batch is done I will start using yeast from my bank.
I plan on using some 1056 for a few batches probably in late July or mid Auguest
 
I've frozen about 30 vials..but have not reconstituted any yet.The vials seems to be sealing properly.

I have some Denny's Favorite 50 on the way that I will be putting into the bank.
Once that batch is done I will start using yeast from my bank.
I plan on using some 1056 for a few batches probably in late July or mid Auguest



Very cool. good to know. I wonder if anyone has done this with cropped yeast. I got quite a bit of cropped British Ale Yeast from the Wynkoop and I am not thinking I am going to use it all in the two batches I am doing tomorrow. He said it was good in the fridge for a week so I want to try and save some of it
 
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