Has anyone testet PrimerBank? - (Apr/20/2006 )
I highly recommend PrimerBank. I've used it for several genes and always had success. The products always resulted in a single band and a clean dissociation plot. (I didn't have any primer dimer problems like someone else mentioned.)
They do not tell you if the primers span exons, so you'd need to check that yourself.
Also, make sure to use the protocol they suggest on their website, not the default settings.
I'm using 16 of their primer pairs (for mouse) and they all work fine.
I had to modify the real-time program because I'm doing the pcr in capillaries and I don't have to wait for 30 sec for elongation, etc. At first I also tried them at 58 C, and they worked at my first trial. Then I tried 60 C (as they said) and they worked again.
Anyway, I strongly recommend it if you don't want to waste time on determining the annealing temperature.
I use RTPrimerDB (Real Time PCR Primer and Probe Database). 95% of the primers i used were VERY good (no dimmers, clean product).
i recommend it .
http://medgen.ugent.be/rtprimerdb/index.php