What type of mutations occur during PCR?
wolfsc at ibms.sinica.edu.tw
Thu Aug 16 20:29:00 EST 2001
There are some protocols that use just Taq and a *standard*
PCR for mutagenesis. Acoording to the literature, Taq almost
only makes substitution errors and is biased towards cratin
bases/ base combinations. If you can get hold of Stratagene's
PCR mutagenesis kit manual (GeneMorph/Mutazyme, it
should be available on their website), you can get some data
on that. Frameshifts and deletions are rather rare. If you want
that, you'll probably have to include intercalating dyes like
EtBr or SYBRgreen in your PCR.
Are you concerned about covering all in sequence space?
Just go ahead! Don't worry too much about that.
> The frequency error-rate when doing PCR with pfuTurbo is
> 1.3x10-6. Does anyone know what kind of mutations is the most
> usual? For example, is frame-shift mutations as usual as
> substitutions? I would guess that it's more frequent to
> experience substitutions than deletions and additions.
> Trond Erik Vee Aune
[tiss mezzage wahs broduceRd using TYPO GENERATOR zoffwer]
Dr. Wolfgang Schechinger
Lab N233 (c/o Dr. Steve Roffler)
Institute of Biomedical Sciences
128 Yen-Chio Yuan Rd. Sec.2
e mail wolfsc at ibms dot sinica dot edu dot tw
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