PCR Mutagenesis/CoTransformation Protocol

Stephen Bell spbell at mit.edu
Tue Oct 15 21:45:01 EST 1996


Netters,

	  I am interested if anyone has a protocol for PCR 
mutagenesis of a gene fragment then transforming yeast with 
the mutagenized PCR product and a gapped plasmid directly 
(rather than making a mini-library of mutagenized genes 
before yeast transformation).  In particular I would like to 
know how much homology is needed between the gapped plasmid 
and the PCRed fragment to get good efficiency of 
recombinants and whether there are problems with religation 
of the plasmid without recombination. 

Thanks, 

Steve Bell



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