DNA extraction protocol for use on Crustaceans..?

TIM CHIPMAN 901106c at axe.acadiau.ca
Thu Jul 1 11:22:29 EST 1993


I'm posting this for a friend, who is trying to use DNA preps from corophium 
volutator, a amphipod. The DNA is for use in RAPD PCR reactions, and is not 
working at all. When his DNA preps are mixed with my honey bee DNA preps 
(Which do work by themselves), the two togother fail to produce any bands. 
Hence, there is something in his DNA which is poisoning the reaction maybe..?

I am wondering if ANYONE has had any luck doing DNA preps like this for use 
with PCR. Our current protocol (in a pillbottle) is:

Freeze, thaw, lyse in SDS buffer, add KOAC, cool, then phenol,
chloroform-isoamyl, chloroform extractions, isopropanol pptn, cold EtOH wash,
wick dry, parafilm overnight, resuspend in TE.

I would greatly appreciate any help!

Thanks!

Tim Chipman
901106c at axe.acadiau.ca



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