yeast plasmid isolation

Varga varga at bart.nl
Tue Jan 22 09:01:19 EST 2002


Dear yeasters,
We have some problems with DNA isolation of yeast. For some reasons, I
obtain little DNA, however, after subsequently transforming E.coli, in hope
to recover our desired plasmid, we never yield colonies on the selectable
carbenicilline (for the plasmid) plates. In addition, we didnot succeed
either with the clontech-kit for yeast plasmid isolation.
My question is: is this a common problem that occurs particularly when
isolating DNA form yeast, and how can we overcome the problems. Is good
enzym treatment better than mechanical disruption of yeast cell walls? We
also tried regular purification of the DNA, however, with no substantial
differences.
Hope you guys can help me out with this.
Thanks so much


George Varga
Keygene. N.V.
the Netherlands
george.varga at keygene.com






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