low Tm primer PCR
steven heckt
shecht at lamar.ColoState.EDU
Tue Dec 15 17:08:09 EST 1992
You should not have to lower the extension temperature. In
working with much lower Tm primers, you can consider the
extension rate of your pol, which is slow but still going
at lower temperatures, and sa you get extension the Tm will
increase. So- you may consider using ramps; ie, ramp down to
annealing slowly to allow the best matches and ramp up to
your extension temp to allow the interplay of annealing and
extension to work itself out.
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