Beta-Actin good RT-PCR Control?

Richard R. Hardy hardy at
Fri Dec 30 20:15:25 EST 1994

In article <smh1008-311294010018 at>,
smh1008 at (David Huen) wrote:

> This might be a bit of a dumb question but it has puzzled me for a long
> time. How do you use beta-actin as an RT-PCR control when genomic DNA has
> beta-actin pseudogenes (it does, doesn't it?)? Surely under these
> circumstances, irrespective of the integrity of your mRNA and success of
> your RT reaction, there should be enough genomic DNA carryover to at least
> occasionally generate a false positive in RT-PCR.
> David Huen aka Puzzled, Dept. of Pathology, Univ. of Cambridge.

Dear Puzzled in Cambridge,
         Well, with our particular b-actin primers (this is in mouse), we
see a band about 70-100 nt larger than the c-DNA band if there is
contaminating DNA - DNA alone gives this band.  We usually don't see it
(we routinely treat our preps with DNAse).

R. Hardy
Member, Institute for Cancer Research,
Fox Chase Cancer Center, Philadelphia, PA
(215) 728-2463

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