I recovered the cultures. All worked.
On Wednesday, February 11, 2015 at 12:05:17 AM UTC-8, Patrik D'haeseleer wrote:
-- To recap, I had cultures with 40%, 20% and 10% glycerol. All were created on 6-25-14 and placed at -20 in a frequently used box.
When I took the cultures out, 40% was completely aqueous. 20% was partially frozen, and 10% was barely frozen (similar to a very fine snow cone). I vortexed them all for a few seconds, which mixed them up a bit. Still, after that there was cells stuck to the bottom. The 40% I was able to mix up more, and after pipetting a few times in and out I got 20µl near the bottom (higher concentration of cells) into a culture tube. I did that with the others, although less efficiently of course because they were frozen.
10% looked to have the highest cell growth rate after, but that could just be an error on my hand on perhaps taking cells too close to the bottom. 20% and 40% were closely matched in their visible RFP expression. All had nice cell growth.
My recommendation then would be 40% for now: the cells have stayed good and it being aqueous allows for you to get cells out quicker and easier. Yes, this is not a very well controlled experiment, but for simply storing cells it might be enough
I'll email you on the DIYbio culture collection! Perhaps we could get something worked out.
-Koeng
On Wednesday, February 11, 2015 at 12:05:17 AM UTC-8, Patrik D'haeseleer wrote:
On Monday, February 9, 2015 at 10:40:56 PM UTC-8, Koeng wrote:I think it would be interesting if biohacker spaces got together and each bought a -20 freezer for redundant storage of cultures from other biohacker labs. Not only would it allow for everyone to keep their cultures safe from a freezer malfunction, but the other labs also get free strains to use in their research. Win-win all aroundAgreed. Counter Culture Labs has a -40 freezer, which could be a great resource for something like this. We also have a centrifugal evaporator system available that we could use to lyophilize strains for room temperature storage and transport. Maybe we should apply for a grant from ASM to set up a little DIYbio culture collection...Patrik
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