Does RNA - PCR ??
Mon Jul 17 04:13:19 EST 1995
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.
Thank you for reading this article and I hope you can help.
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