Freezing yeast cells - (May/21/2014 )

Good morning everyone,

I´m actually using Saccharomyces cerevisiae yeast cells (AH109), and to store them at - 80ºC, I usually use 12% of DMSO (like I do for bacteria: E. coli and Caulobacter crescentus), but few weeks ago some peoples told me that it is better to store yeast cells with glycerol and not DMSO, is it true ? If yes, which concentration of glycerol should I use ? I also want to say that when I thaw the yeast cells (after a freezing with DMSO), the cells have a lot of difficulties to grow (approximately 48h to reach an OD of 1 in rich medium ), do you think that it is due to the freezing with DMSO ?

Thank you   

microbiologiste24 on Wed May 21 06:46:56 2014 said:

Good morning everyone,

 

I´m actually using Saccharomyces cerevisiae yeast cells (AH109), and to store them at - 80ºC, I usually use 12% of DMSO (like I do for bacteria: E. coli and Caulobacter crescentus), but few weeks ago some peoples told me that it is better to store yeast cells with glycerol and not DMSO, is it true ? If yes, which concentration of glycerol should I use ? I also want to say that when I thaw the yeast cells (after a freezing with DMSO), the cells have a lot of difficulties to grow (approximately 48h to reach an OD of 1 in rich medium ), do you think that it is due to the freezing with DMSO ?

 

Thank you   

I can't say what the difference is to be honest: I never used DMSO for yeast cells, but I use whats called stock medium.

Its 2 grams of bactopeptone, 1 gram of yeast extract and 30 ml of 87% glycerol. This is in an end volume of 100ml.

I use this to store yeast cells.

Its also normal that they grow a bit slower after being frozen, but 48 hours seems a pretty long time to reach an OD of 1. (Altough, no idea how much you add of the cells to how much volume...)

Thank you for your reply Pito

Hey,

I have always used glycerol. Currenctly I am using this procedure http://openwetware.org/wiki/McClean:_Glycerol_stocks_%28yeast%29

Best of luck!

Yeah I do use glycerol .

it preserves the structure of the cell better. I believe I use a 30% of glycerol. Pour 1mL of it and 1mL of the yeast broth so the final concentration would be 15%.