DEPC in RACE?
hxm8 at po.CWRU.Edu
Thu Aug 18 16:41:45 EST 1994
Well, I'm about to try a 5' RACE for the first time. Current
Protocols calls for a 3 hour incubation for the the primer/RNA
annealing step. My question is.... Do I use DEPC-treated reagents
at this step? Three hours is a LONG time, and I'm paranoid about
leaving my RNA at 37 degrees for that long, even WITH DEPC. But,
I'm also well aware that DEPC can kill Taq activity. This leaves
me at a loss for what to do. Will a subsequent precipitation step
get rid of enough of the DEPC to allow the Taq to work?
Thanks for any help by you RACE experts.
Harry Menegay hxm8 at po.cwru.edu
Department of Neuroscience
Case Western Reserve University
Cleveland Ohio, 44106
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