Degenerate PCR: concentration of primers?

J.B. Herrick jherrick at lanl.gov
Wed Nov 5 18:12:31 EST 1997


I routinely work with degenerate primers and have often found that a 10X
concentration of primers is needed for efficient amplification. As
others have pointed out, if the PCR is simply not working at all, it's
best to just order some new primers. However, if you're getting the
correct fragment amplified, just not much of it, I would recommend
optimizing the primer concentration. Other helps are the use of DMSO
(sometimes it helps, sometimes it doesn't) and especially the use of
'Touchdown' PCR.

Good luck.



Jim Herrick

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James B. Herrick, Ph.D.
Microbial Ecology and Genetics
Environmental Research and Development Group
University of California/Los Alamos National Laboratory

Mail to:
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Los Alamos National Laboratory
Los Alamos, NM 87545

jherrick at lanl.gov
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