Hi all,
It is about running the multiplex PCR. When I did the PCR separately, the band can be run individually. However, when I mix two primers set together, and half the total vol.. I cannot see any bands in the gel. Could you please offer any suggestions so that to improve? Thank you. 
Did your primers self-binding together? (Primer Dimer)
Tell us more of your protocol so that we can help you more...
I got the primers, 402bp (Tm: 66-77C), 308bp (Tm: 76C) & 245bp (Tm: 77C). with annealing 58C for 30s (according to protocol).
If I decrease the annealing temp, how low should i adjust?
if i decrease PCR primer & increase the template? but they work when do it in separate.
what about increase MgCl?
Thank you very much in advance.
Edited by icarus.rachel, 01 December 2009 - 07:58 PM.