I'm fresh in running PCR. I have an enquiry about the PCR cycling program on detection genus of Chlamydia.
I'm using the primers stated its Tm is 56-58C.
however, the PCR cycling conditions are as follows
1. 10min at 94C
2. 15sec at 95C for 40cycles
3. 15sec at 55C
4. 30sec at 72C
I have no product for this conditions at the end. I'm thinking to do a gradient to measure the optimum temp for annealing
Please advice for other options. Thank you very much.
Edited by icarus.rachel, 09 March 2010 - 08:32 PM.