Freezing of S. cereviceae competent cells

George Theodoris ez003450 at boris.ucdavis.edu
Wed Aug 14 13:34:37 EST 1996


Botma Visser (pbv at rs.uovs.ac.za) wrote:
: Hi
: 
: I am busy transforming S cereviceae using the Li-Ac method. 
:  Is there any way to freeze the rest of the competent cells 
: away for later use as with E. coli?  

I make the up the TE/Li-Ac buffer with a final concentration of 20%
glycerol and use this for all my yeast transformations. This way I don't
have to worry about adding glycerol or DMSO to the competent cells before
freezing, and the Lithium concentration is not diluted by the pre-freezing
addition of glycerol. If I want to freeze away cells I just aliquat
200 microliters into eppindorf tubes and freeze them at -70,  after 
re-suspension in the Lithium/TE/glycerol buffer and before addition of
DNA.
-George



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