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
