pcr problem with gc rich sequences

Nick Jacobsen jacobsen at cf.ac.uk
Tue Jun 4 04:20:02 EST 1996

I had similar problems with a very GC rich sequence. I solved the problem 
by first designing primers with high Tms (about 80 C). I then used NEB 
Deep Vent exo- (not designed for PCR but works anyway). The reaction 
conditions were 98 C 5mins, (72 C 1min,98 C 1min)X30. This amplified the 
desired product from a gene that previously didn't want to know.

hope this helps


More information about the Methods mailing list