HELP ... yeast transformations

Hoffmacs Hoffmacs at HERMES.BC.EDU
Tue Jul 26 13:47:55 EST 1994


Dear Julie et al.,
We are working with strains in S. pombe that do not respond well to standard
transformation protocols.   We have therefore tried them all.  One of the ones
that made a big difference published in BioTechniques by Randy Elble (1992, I
believe).  It involves an avernight incubation with the PEG/LiOAC solution and
no heat shock.  I would recommend it if you are having problems with the more
commonly used procedures.
Charlie Hoffman
"Let's run it over a column and see who elutes."
________________________________________________________
To: yeast at net.bio.net
From: Julie on Mon, Jul 25, 1994 5:31 AM
Subject: HELP ... yeast transformations
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From: Julie <JANDREWS at fs1.pa.man.ac.uk>
Subject: HELP ... yeast transformations
Date: 25 Jul 1994 09:59:11 GMT
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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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