Problem: removal of DNA from RT-PCR using DNase

teresa_mogul at my-deja.com teresa_mogul at my-deja.com
Mon Sep 11 20:37:00 EST 2000


Hi

I have just begun RT-PCR experiments. They are going very well so far
with beautiful amplification as show on agarose gel. However, when I run
the no-RT control I have a very faint band indicating that I have not
fully removed the DNA from my RNA. I measured the amount of RNA/DNA in
my sample and according to the manifacturers notes 30 minutes at 37°C
with 1U DNase should be ample to remove my DNA. Increasing the time does
not seem to help. Has anyone got any suggestions? I am using glass
microfibre plates and guanidinium thiosyanate for extraction could there
be a DNase inhibitor in my DNA/RNA extract?

Thanks in advance for any help
Teresa


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