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
