Taqman Assay Question
dknorr at lynxgen.com
Wed Mar 1 15:35:56 EST 2000
> Jeff Fairman wondered about optimizing TaqMan reactions...
PE also has a pretty good piece on their website about optimization.
Briefly, their philosophy is to give the enzyme everything it needs to be a
good polymerase, and then change the primer sequence and concentrations to
optimize the conditions. The most common approach to PCR optimization seems
to be optimizing the reagents and cycling conditions to the primer. By
holding the reagents and cycling constant, any assay can be run with the
same mix, and assays can be run side-by-side. The optimization for any
TaqMan system take only a couple of runs. True, you might find the overall
performance of that assay to be less than you'd hoped for, but it will be
optimized. You'll have to develop another primer/probe set that you hope
will be better.
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