Does RNA - PCR ??
Andy Law Big Nose
Andy.Law at bbsrc.ac.uk
Mon Jul 17 12:17:42 EST 1995
In article <DBusA7.Lv5 at info.swan.ac.uk>, BABAKER at Swansea.ac.uk wrote:
> Does RNA PCR??
> I am doing RT-PCR and get positive results even when just PCRing RNA
> using cDNA primers and Taq DNA Polymerase, with no cDNA or genomic DNA
> anywhere close by!!
> Is it possible that the gene sequence of the mRNA which would only
> contain 1 or 2 U's instead of the T's in the primers sequence would be
> amplifiable by the cDNA primers? As annealing would probably jump the
> U's in the mRNA for that gene and then continue in the PCR to give
> the correct sized band?
> I dont think that DNA is contaminating my RNA prep as I use the
> Acid-Guanidinium thiocyanate method of isolation, which should
> erradicate the DNA.
My first inclination would be to test the primers and the Taq for
contamination. I assume that you include blanks in your PCRs?
( Andy.Law @ bbsrc.ac.uk )
( Big Nose in Edinburgh )
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