5' RACE

Malcolm Moos Jr., M.D., Ph.D. moos at helix.nih.gov
Fri Oct 7 09:14:54 EST 1994


We have found the Clontech Kit to work reliably when the RNA is good.
We have had the best luck with random primed cDNA synthesis and RNase
H- MMV RT in the presence of 5 mM methyl mercury, which is a departure
from the manufacturer's protocol. We have done several side by side
comparisons (~6) of random vs gene specific cDNA synthesis priming, and
the former has always been better.

Malcolm Moos Jr., M.D., Ph.D.



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