PCR Design

[Brian Ballard]

> Only rarely can a poor primer be rescued by changing the
> annealing temperature. 

I absolutely disagree with this statement. I find that changing the 
annealing temperature is often the best way of 'rescuing a "poor" 
primer'. I frequently use the Biometra T-Gradient block for finding the 
optimal Tm of a primer pair; especially when I am forced to design 
'poor' primers due to the nature of the experiment. You'd be amazed at 
how often the optimum temperature is nowhere near the Ta you'd predict 
on theoretical grounds.

Brian