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Freezing yeast cells

yeast freezing dmso glycerol

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#1 microbiologiste24

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Posted 20 May 2014 - 10:46 PM

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 sad.png ), do you think that it is due to the freezing with DMSO ?

 

Thank you smile.png  



#2 pito

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Posted 21 May 2014 - 01:47 AM

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 sad.png ), do you think that it is due to the freezing with DMSO ?

 

Thank you smile.png  

 

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...)


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#3 microbiologiste24

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Posted 21 May 2014 - 05:45 AM

Thank you for your reply Pito smile.png



#4 Alfahelix

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Posted 07 July 2014 - 03:38 AM

Hey,

I have always used glycerol. Currenctly I am using this procedure http://openwetware.o..._stocks_(yeast)

 

Best of luck!



#5 Adriana Reis

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Posted 01 August 2014 - 03:21 AM

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%.







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