help: long pcr problem (urgent)

Parent, Eric 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
small 
> 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. 


Hi Chris

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.

Eric Parent
Maurice Lamontagne Institute
Fisheries and Oceans Canada
 Invertebrates and Experimental Biology
850 route de la mer
Mon-Joli, Qc
Canada
G5H 3Z4

parente at dfo-mpo.gc.ca
> http://www.qc.dfo-mpo.gc.ca/iml/fr/intro.htm
> 
> 
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