HELP ... yeast transformations

Julie JANDREWS at fs1.pa.man.ac.uk
Mon Jul 25 04:59:11 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.

Thanks in anticipation



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