PCR elongation time?

Chris LaRosa clarosa at biocomp.unl.edu
Mon Mar 20 13:15:57 EST 2000



Sophie Launey wrote:
> 
> Hello everybody,
> 
> My question is: how short can the elongation time in a PCR cycle be?
> (for fragments of less than 300 bp)

Rule of thumb at 72 degrees with Taq polymerase is 1 min per 1000
bases.  Perhaps short as 20 seconds for you?
> 
> The side question is: can I hope to reduce band fuzziness by shortening
> the PCR steps. I'm doing PCR (of microsatellite loci) with incorporation
> of some fluorescent dCTP and get very fuzzy bands and lots of
>

Microsatellites can be tricky if there are dinucleotide repeat regions,
causing "stutter"




More information about the Methods mailing list