site-directed mutagenesis problems
wsun at jeeves.ucsd.edu
Wed Dec 9 16:15:26 EST 1992
I recently posted the problems I had and appreciate your
responses. However, I am still not getting the mutants using
the Bio-rad kit. I am getting transformants with the negative
control DNA - where I make the second strand in the absense of
any mutagenic primer.
Just recently, I found that my SS DNA was able to transform XL1
blue competents. Apparently the SS DNA prep was contaminated
with DS DNA. Has anyone had experienced this problem? I am
making SS DNA from the Bluescript plasmid in CJ236 cells. I am
thinking of gel purifying the SS DNA. Is this a logical or a
stupid idea? I appreciate any comments.
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