PCR help with B. cepacia


Tue Apr 3 07:12:34 EST 2001


I'm a final year degree student working on a project to identify epidemic
strains of B. cepacia using Cbl-A and BCESM markers by PCR. I've obtained
amplicons for the Cbl-A fragment but can't seem to get any for the BCESM
fragment. I know that Taq polymerase is working as I've been using it for
Cbl-A and the positive control(uni5/uni6) has showed amplicons for Cbl-A but
not BCESM. I've also increased the cycle number from 25 to 35 and annealing
temperatures of 63, 55, and 58(all recommended temperatures in various
papers related to this experiment) degrees have been used with no success.
 Could anybody suggest anything else that could be interfering with the
reaction
and preventing amplification of the BCESM fragment. Any help would be much
appreciated.

thanks
sharon







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