I've actually experimented on this :D
On Monday, February 9, 2015 at 8:57:58 PM UTC-8, Patrik D'haeseleer wrote:
-- https://groups.google.com/forum/#!searchin/diybio/4$20months/diybio/u4mxuMNTyMw/VbgwOQA5IUwJ
That was a couple of months ago, so I'll check it again if you want me to see. Essentially, I tried it in -20 at 10%, 20% and 40% glycerol and tried to recover cells from that. They all worked. It seems like storing cells is pretty robust even in these conditions (for 4 months, so far)
I usually just put in 80% glycerol at half the freezing stock (250µl of 80% glycerol for 500µl of cells) and that's worked. To my knowledge, I'm the only one who has done a controlled experiment on this. It'd be great if someone else could put up their research or start their own, I would like this to be reproducible so I can recommend it! I'm working on a standard part distribution for DIYbiologists and it'd be great to have -20 storage protcols
(It also should be noted that I used E coli. S cerevisiae may be different because yeast freeze differently and are more prone to lysis (they just die more in stocks) and B subtilis doesn't even need freezing storage. I store B subtilis on paper)
-Koeng
On Monday, February 9, 2015 at 8:57:58 PM UTC-8, Patrik D'haeseleer wrote:
Hi all,The gold standard for long-term storage of strains is glycerol stocks in a -80 freezer. Or even an ultra-low at -150C, or liquid nitrogen at -196C.However given that most DIYbio labs don't have access to those kinds of resources - what are the best recommendations for storage in a -20C freezer?50% glycerol? 15% glycerol? 10% skim milk? Stab cultures?Opinions / best practices wanted!Patrik
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