pcr and mutant alleles

Joel A. Kreps jak8 at po.CWRU.Edu
Thu Jun 4 11:32:45 EST 1992

Dear Network,
I would like to use PCR to specifically amplify the sequence of a
mutant allele of a duplicated gene to identify the mutation.  I
can isolate poly A RNA which contains 20X higher levels of my 
mutant allele relative to the wild-type isozyme.  I know the seq
of the wild-type of both isozymes and can generate isozyme spec
ific primers.  I plan on amplifying from cDNA, for a full length
cDNA the PCR product would be 1800 bp.  I would appreciate any 
helpful hints from people who have done similar things as to the`
best way (ie which polymerases) to amplify and then the best way
to go about sequencing.

Joel A. Kreps

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