HELP.... Yeast Transformations

Tony ajd at dir.mcc.ac.uk
Fri Jul 22 06:10:49 EST 1994


we are currently using the Lithium acetate method to transform an 
E.coli shuttle plasmid into S.cerevisiae. However, contrary to what 
we would expect, we find that we do not see any improvement in the 
transformation efficiency when we include a heat shock of 42 degrees 
for 15 minutes. We are finding that with or without the heat shock we 
routinely get about 200 transformants from 1ug of DNA added to 40ul 
of competent cells. We expected to see about a tenfold improvement 
with the heat shock step included.

Has anyone out there had a similar experience, or could you perhaps 
comment on why we are seeing this result. All responses would be 
gratefully received as we need to attain a higher transformation 
efficiency.

Thanks in anticipation

Tony D'Emanuele (ajd at dir.mcc.ac.uk)



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