FW: Mismatched primers

Deanne Bell dbell at fresno.ars.usda.gov
Wed Oct 2 17:25:17 EST 2002

:after hotstart (because after hotstart there will be no more chance
:for the harpin to form if you have stringent annealing conditions).

what if the hairpin / primer dimer "forms" before the 25bp primer anneals?

:Second: Get yourself MeltCalc anyway (http:www.meltcalc.com). This
:software is freeware for non-commercial use. It calculates exact
:primer Tms even considering mismatches. For description see Schütz and

I find most of these Tm claculators to be grossly overestimated.  They may give a theoretical Tm, but in actuality the recommended temps are not optimal.

Deanne Bell

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