yeastplasmid rescue

Hoffmacs Hoffmacs at HERMES.BC.EDU
Mon Mar 14 10:36:07 EST 1994


We use Smash and Grab preps which are extremely fast (about ten minutes for
transformations, about 45 minutes for Southern quality DNA).  See Gene 57
(1987) 267-272 for the protocol or Current Protocols in Molecular Biology (Unit
13.11-Supplement 24).  One thing we worry about in S. pombe is integration of
the plasmid.  Are you certain your plasmid is still autonomous?  If not, you
can still get it by isolating genomic DNA (also by Smash and Grab), digesting
with an appropriate restriction enzyme, ligating and transforming E. coli. 
Good luck.
Charlie Hoffman
Boston College
"I'd rather be fission."
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From: djdlab at bimcore.cc.emory.edu on Sun, Mar 13, 1994 4:45 PM
Subject: yeastplasmid rescue
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From: ckrudy at bimcore.cc.emory.edu (Margaret M Lanterman)
Subject: yeastplasmid rescue
Date: 13 Mar 1994 21:33:17 GMT
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I am trying to recover a cen plasid from yeast and transform E. Coli.  I have
tried a boiling prep protocol that didn't work for me.  Does anyone have a
protocol that always works?  Does the strain of Ecoli matter- I used HB101.
I really need this to work ASAP help!!!!
Margaret M Lanterman








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