Yeast maxi plasmid preps?
Kathy Baynton
baynton at esbs.u-strasbg.fr
Fri Mar 7 06:42:37 EST 1997
Dear yeast-handlers,
I was wondering if anyone knows if it is possible to produce maxi
preps of plasmids from yeast that are relatively clean and in
respectable quantities (ie. using 2mu-based origin plasmids) as is
done in E. coli? (ie CsCl purification, Wizard). I am already aware
of plasmid isolation preps from yeast (Hoffman and Winston I believe
is the reference), but these preps are pretty dirty, the plasmid is
not really "purified" and we always end up having to re-transform into
E. coli in order to recover plasmids that are "clean enough" to do
sequencing, cloning etc. I am also aware of PCR-based sequening that
can be done on yeast-isolated plasmids but for what we're thinking of
doing, I'm not sure that this would be an appropriate method.
Any information, advice...anything...would be tremendously
appreciated!! Thanks!!
Kathy
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