Site directed mutations

Jeffrey Alan Silverman jeffs at elsie.nci.nih.gov
Tue Apr 20 07:19:53 EST 1993


Has anyone out there any experience with Enzymatic Inverse PCR?  I am trying it
with little success.  After making the primers - exact match for 20 bases containg a restriction site for a class !! enzyme- I get NO product in the pcr 
reaction!! (despite trying the obvious manipulations in temperature, Mg, various
times of anneal and extensions etc).... So does anyone actually use this method?
how did you get it to work? 
Related: if you don't use this method but rather favor another for site
directed mutagenesis... I'd be interested in hearing about that too...
Thanks  ;-)
Jeff



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