PCR stutters through oligo-A regions
pnorton at lac.jci.tju.edu
Wed Mar 20 17:32:59 EST 1996
In article <4in1cj$nu6 at majestix.uni-muenster.de>, Juergen Brosius
<RNA.world at uni-muenster.de> wrote:
> Thanks for the idea. However, we had a similar problem in cDNA clones
> containing a stretch of 12 C-residues. In this case we do not know
> whether the stuttering or slippage was due to reverse transcriptase
> (first strand) or DNA polymerase (second strand cDNA synthesis) or both.
Reverse transcriptase is a likely culprit, it has been reported to
stutter at A tracts (U tracts on template RNA), and I have seen this
myself in primer extension reactions.
Juergen, it wasn't clear in your original post if you were performing
an RT step prior to the PCR reactions that are giving you trouble, or
whether the starting template is DNA. Which is it?
Pamela A. Norton, Ph.D. Assistant Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107 p_norton at lac.jci.tju.edu
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