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Display PCR help

Gene rxr generxr at aol.com
Wed Jul 20 22:28:01 EST 1994


We had some promising and reproducible results coming from human tissues
and the GenHunter kit until 6 weeks ago when much of our efforts "went
down". Reading the Biotechniques article (different tubes -> different
results) we have tried various tubes w/o success. Also unsuccessful have
been: changing RT, changing label, new kit, changing taq source, new RNA
extraction, different gels, different cycler, differing annealing temps,
different primer sets, etc etc. Most frustrating is the randomness of
outcomes. We often have gels in which one of 8 RNA samples give nice band
patterns, but all others for that day are blank. (No bands whatsoever, or
very weak ones that don't get much better on prolonged exposure) At first,
GenHunter said it must be our fault, but more recently they seem to be
wondering themselves. On many occasions, we have been unable to get even
their control RNA to work. Is anyone out there having similar problems?
Any ideas?



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