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Primer concentrations with rapid PCR

nvahsen at hot.hgmp.mrc.ac.uk nvahsen at hot.hgmp.mrc.ac.uk
Mon Jan 3 14:53:50 EST 2000


Hi Lars,

I think you are right.
For a 20mer primer at 0.2 M Na-equivalents you can calculate a t1/2 of
approx. 3 sec. for annealing. And annealing time is should be short on
the LightCycler. More than 10 sec. makes no sense.
Read Wetmur JG, Crit Rev Biochem Mol Biol 1991, 26:227 for more.

Best regards
Nicolas





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