FW: Mismatched primers

[Deanne Bell]

: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.



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Deanne Bell
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