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

( Andy.Law @ bbsrc.ac.uk )
( Big Nose in Edinburgh )



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