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Re Display PCR help

Steve C schamberlain at molbio.uoregon.edu
Fri Jul 22 10:14:22 EST 1994


I have been doing this method for about 4 months now and have had mixed
success.  Sometimes I get ladders, other times not and after many control
experiemnts to nail down the offending step(s) I have little to report.  I
DO know that high quality of the input RNA is absolutely essential to the
success of the assay.  Try Operon's primers, they're 1/30th the cost of
GenHunter's and "work" just as well.  I feel the first two or three cycles
are the most critical in the PCR step and have increased the denaturing
time to 2 minutes prior to the first annealing and extension.

Steve



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