PCR with primers of different lengths

Luc Simon Lsimon at alnus.for.ulaval.ca
Fri Jan 10 11:38:56 EST 1992


 Ethan Strauss writes:
 
> I am trying to add approx. 60 bp to the 5 prime end of a piece of DNA. Is it
> possible to do PCR with one primer of 80 bp (20 overlap and 60 new) and
> another of 20 bp (all overlapping). I suspect that their may be problems
> with melting after the first few rounds of PCR. Does anyone have any experience
> with this kind of thing?

There shouldn't be any problems, it has been done by Myers et al. with 
their GC-clamps. See Chapter 7 in PCR technology, H.A.Erlich ed., Stockton
press.1989



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