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Old 10-27-2011, 01:32 PM   #41
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Wow, 2 weeks in a defrost freezer? I would assume adding freezer packs for stabilizing would help for longer term storage, but that is still interesting to know.

My thoughts are that the day in the fridge will help with glycogen build up storage in the cell, and not glycerol diffusion, that should happen rather quickly.

This is good stuff. Should drive the point home how easy it is to make your own ready for starter yeast library. I've been using vials, maybe 3/4 inch compact slurry in the bottom (2-3 mls worth) and stepping up starting at 200ml. They're ready in 1-1.5 L within 48 hours.

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Old 10-27-2011, 06:37 PM   #42
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Was only two weeks because I got impatient (that and we were limited on freezer space :lol ) and I've been toying with the process. Additional to those was a vial I placed in some steaks I have in the freezer with the same mix but a lot less cells and I anticipate very VERY long term storage out of it. Once I have a bit more freezer space to work with I'm going to pick up one of those insulated lunchboxes and go the frozen coldpack route. I can't speak for the super-longterm currently but there are enough people above saying it works pretty well.

It does seem to make a very noticeable difference to at the very least give it some fridge time before freezing it. I wonder, however, if you were to chill it in ice water (or just ice) until super cold before freezing it if you'd get the same results or if it's the time the yeast has to either build up glycerine inside the cell and/or it's glycogen stores as you mention above.

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Old 10-27-2011, 06:41 PM   #43
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Oh, and as a sidenote I did have a brain fart previously a couple years ago and placed my washed yeast in the freezer and it seemed to have killed the yeast quite efficiently (all slurry combined after thaw and no activity in 5 days in a starter). And it was only in there for two days before I realized my stupidity and took it out. If any did manage to survive it couldn't have been that much.

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Old 10-27-2011, 07:33 PM   #44
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Quote:
Originally Posted by Accidic View Post
Oh, and as a sidenote I did have a brain fart previously a couple years ago and placed my washed yeast in the freezer and it seemed to have killed the yeast quite efficiently (all slurry combined after thaw and no activity in 5 days in a starter). And it was only in there for two days before I realized my stupidity and took it out. If any did manage to survive it couldn't have been that much.
Well, I guess you just proved the importance of glycerine, in case anyone asks! Again, it's interesting to know the limits of these things. If I had to guess, I would have thought there would be at least some fraction of superhero cells that made it through the freezing process. Though freeze/thaw cycles are a normal cell lysis protocol for cell work, so I guess there's some validity there.
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Old 10-28-2011, 03:42 AM   #45
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seriously awesome! this is exactly how we will have to do things here in El salvador with our micro brewery. We gotta be yeast farmers!

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Old 10-28-2011, 03:14 PM   #46
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There may have been a small number of cells but in the timeframe I gave them they did not multiply enough to have visible viability. I had an old wl vial that had gotten packed (forgot I had it honestly) when I escaped Cali and shipped ups in the summer. High heat + ~20 month old vial behaved the same way. There were probably some living cells and maybe I could have nursed enough back to use it but made more sense to just but another vial. If I couldn't have gotten the strain again tho I probably would have pushed harder.

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Old 11-04-2011, 09:05 AM   #47
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This one of the most fascinating threads I've ever seen here! +1
Two questions:
1 We seem to be referring interchangeably to glycerine and glycerol. I just wanted to know if it really is the same product, and if any kind of glycerine would do, or do you need "Food grade" stuff.
2. Has anyone tried this with any other anti-freezing agent? Agar, Gelatine, Alcohol? Rockey-Road ice cream? I'm just curious.

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Old 11-04-2011, 09:19 AM   #48
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Glycerin, Glycerine, Glycerol = same stuff. I would use pharmaceutical grade because it will be sterile when you buy it. I don't know what the price is in the drug store but it should be readily available. If you don't see it ask the pharmacist. You can save on the volume if you let your yeast settle out first and pour off the wort. Only about 5% of the total volume of the starter is yeast. Bring that up in a 15-30% solution made up in boiled water and you should be fine. Doing all this at refrigerator temps should work well.

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Old 11-04-2011, 11:01 AM   #49
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Seeing as right now my kitchen feels about as cold as the inside of my fridge that shouldn't be a problem. Thanks for the info. What about using other stuff? Anyone try?

Also, if I can't get my hands of sterile glycerin, can I sterilize it just by boiling the mixture, or would that ruin it somehow?

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Old 11-04-2011, 02:41 PM   #50
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You can sterilize it in a pressure cooker just like wort. I use USP grade, but drug store glycerine works just fine. I'll dilute it with water to about 40%, sterilize it in a pressure cooker, store and dilute that 1:1 with yeast slurry before cooling/freezing. Never tried agar, etc, but you really need a cryoprotector, something that penetrates the cell to allow it to stretch and not fracture while freezing.

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