PCR Nightmare

ELADER at AMBION.COM ELADER at AMBION.COM
Fri Feb 12 21:11:10 EST 1999


In article <36C4591C.560E2494 at utmb.edu>,
  Leslie Hutton <lahutton at utmb.edu> wrote:
> I have been haveing problems with background in my RT-PCR reactions. I
> do not see bands when the RT or RNA is left out of my reaction. However,
> if I add RNA which has not been reverse transcribed to the reaction I
> see bands. DNase I digestion of the sample does not eliminate this
> problem nor does it occur with all of my primer sets. Has this ever
> happened to anyone else? What is the cause and how do I eliminate it?
> DNase I digestion does not help. \

This is interesting. A no-rt control being positive with your rna would
suggest DNA contamination. But you say that if you leave out the RT, then you
do not get bands. Then you say that if you add RNA which has not been reverse
transcribed, then you see bands. Which is it? E-mail me and let me help.

Eric, the RT-PCR guy at Ambion.

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