ligation-mediated PCR

John Watson ajwatson at romeo.caltech.edu
Tue Dec 20 15:01:31 EST 1994


In article <3cknlu$cgk at news.cc.utah.edu>, Tom Doak
<tom_doak at hlthsci.med.utah.edu> wrote:
> 
> We are preparing to begin ligation-mediated PCR (LM-PCR), but our reading
> indicates considerable confusion about choices of DNA polymerases (with
> and without 3', and even 5' exonuclease activities), and about the design
> of primers (biotinylated or not, P3 overlapping P2, etc), and a variety
> of other details.  We're seeking up-to-date protocols and literature
> references.  Any suggestions, comments, hints, or gossip would be very
> much appreciated.
> 

When I was doing LM-PCR a few years ago, I used the original Mueller and
Wold protocol (Taq polymerase, increasing Tm's of nested primers, P3
end-labelled with P32) with no modifications.  Can't remember offhand baout
overlapping primers, but I think P# overlapped P2.  Upshot was -- it worked
beautifully.  I can mail you reprint if you want, or just the reference.

john  



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