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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