Function of dUTP in real-time PCR?

Paul Wary paul_wary at yahoo.com
Fri Oct 29 09:43:03 EST 2004


Tom Knight wrote:
> paul_wary at yahoo.com (Paul Wary) writes:
> > I read in a description of mixes used for real-time PCR that they
> > contain dUTP "to prevent re-amplification of carryover PCR products".
> > Can anyone tell me how this works or give me a reference?
> 
> The mix also contains an enzyme which selectively degrades dUTP
> containing DNA.  This enzyme is degraded early in the PCR process.
> Carryover PCR product will have dUTP and will be degraded.  Real
> template will have dTTP, and will not be degraded.  The final
> amplification product will contain dUTP, and will be a poor template
> for the next PCR reaction.

Ummm, where do I actually get any carryover PCR product from? I use
real-time PCR for quantification of gene expression levels, so I
reverse transcribe mRNA to cDNA and use that cDNA for the real-time
PCR. I can't see where I get that carryover PCR product from.

Paul



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