help: long pcr problem (urgent)
ParentE at dfo-mpo.gc.ca
Fri Mar 31 09:06:03 EST 2000
> I'm using Boehringer Expand Long Template. I've been able to amplify a
> fragment, reverse the primers and try to amplify the mtDNA.
Check your GC content of related organisms. In some weird slime molds,
the AT richness forces you to run the pcr at 60 C with 8 min extention.
How pure is your mitocondrial template.
I've sequenced a part of the mtDNA sequence to desing new primers, and it
seems to be fairly AT rich. I didn't purified the mtDNA, just a quick total
DNA extraction. It worked fine for fish.
Maurice Lamontagne Institute
Fisheries and Oceans Canada
Invertebrates and Experimental Biology
850 route de la mer
parente at dfo-mpo.gc.ca
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