Information about the use of DNA diluted in Real Time PCR - (Sep/19/2013 )

Hi,

For the removal os PCR inhibitors for Real Time Detection we should dilute the DNA sample 1:10 fold, consequently we are reducing the amount of DNA available for amplification, this fact cannot give false- negative results?

Thanks

-rmmoniz-

Why not?

-bob1-

So for PCR inhibitors removal, what do you recomend to use in daily diagnosis to prevent false negative results?

-rmmoniz-

There isn't anything that I am aware of. The false negatives won't be entirely removed by diluting 1:10, you could also try 1:100, 1:1000 etc.

-bob1-

But the dilutions that you recommend wont increase the chances of the sample give a false negative result? Mainly in the cases of very minimum loads of DNA/ RNA target?

-rmmoniz-

Potentially, though if you are doing qPCR with good primers, then the reaction should be close to 100% efficient, so detection should work from very low abundance targets.

-bob1-

Ok, thank you very much

-rmmoniz-