HELP. Multiplex RT-PCR with GAP as a housekeeping gene.

Thomas Mohr thomas.mohr at
Wed Feb 14 08:15:56 EST 2001

Dear Colleagues,

currently we're trying to establish a multiplex RT-PCR with GAP as a
houskeeping gene. Intended is the useage GAP primers plus a second
primer pair (and maybe third). However, our GAP products require only
22-25 cycles, compared to 30 to 35 cycles of the others. Coamplification
with GAP primers added after 10-15 cycles lead so far to an almost
complete quenching of the other product. 

Does anybody know of GAP primers fulfilling following condditions:

Annealing temperature 55-60C, required cycles: 30-35 ?

Thanks for your help !


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