PCR Optimization question - (Nov/03/2006 )
How important is it to minimize primer-dimer formation in a reaction? seems like you can drive yourself crazy adjusting Mg and primer concentrations and annealing temps...as long as you get good amplification of your target I don't see what difference it makes if you have some non-specific product formation
-BioDude1-
How important it is depends on the application. For most cases I've encountered, the presence of primer-dimers is unimportant.
-woodshedder-
I generally don't get overly excited about dimers unless I'm doing realtime
-aimikins-
primer dimers - unimportant, unless it is real time or the product is small ~ 100bp and below
-perneseblue-
QUOTE (BioDude1 @ Nov 3 2006, 02:30 PM)
How important is it to minimize primer-dimer formation in a reaction? seems like you can drive yourself crazy adjusting Mg and primer concentrations and annealing temps...as long as you get good amplification of your target I don't see what difference it makes if you have some non-specific product formation
Aside from the problems assiciated with PD, the only really good way to get rid of PD is via careful primer design. I've never had luck with reducing PD by doing those other thing.
Matt
Academic Blog
-Peromyscus-