PCR errors:How many really?
tedm at darkwing.uoregon.edu
Wed May 31 08:15:07 EST 1995
In article <3q81hq$h9d at nntp3.u.washington.edu>, yacman at homer26 (L.
> Okay, I know papers have been rejected because people have cloned using
> PCR and then either 1) not sequenced the clones, and/or 2)not tested
> independently isolated clones for whatever functional assay they are
> interested in. The usual line is that using PCR you take the risk of
> introducing PCR errors into the product. I seem to recall that Taq
> polymerase does have a relatively high error rate, about one in ten
> thousand bases. For most applications, I think this error rate is really
> quite low. Furthermore, anyone who has done any sequencing knows that
> standard sequencing errors exceed (easily) greater than one in ten
> thousand. So, my questions: Anyone care to comment on the 1 in ten
> thousand rate? In my, all-be-it, limited PCR experience I have never
> seen a genuine, confirmed, PCR error. Am I lucky or what? Is template
> selection (e.g. GC content) or amount likely to affect error rates. My
> preference is hearing from voices of experience, rather than theoretical
> gobbeldy gook (sp?)
> Scot Bastian Ph.D.
Refer to a discussion of Taq fidelity by T. Kunkle in PCR Methods journal.
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