primer design for stratagene's quick change mutagenesis

Mark Bowen mb at laplace.csb.yale.edu
Mon Aug 23 08:50:56 EST 1999


I've used this strategy many times to make mutations.  The primer is in
excess over the template so there will always be some primer annealing.
With respect to spurious mutations, I use pfu turbo which has proof
reading capability and have never had a problem with mutation.  I've
done this on plasmids as large as 10 Kb.  If you are really concerned
about it, just subclone a smaller fragment containing your mutation back
into your starting plasmid.

--
Mark Bowen
HHMI/Yale University
Dept. of Molecular Biophysics and Biochemistry





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