question regarding RACE kits

ajwatson ajwatson at romeo.caltech.edu
Tue Apr 12 14:15:25 EST 1994


In article <2nsr09$i0i at news.iastate.edu>
axjl at iastate.edu (Feng Jin) writes:

> I am studying a cuticular wax gene in maize. The transcript is about >1.4-kb, however my largest cDNA is 0.8-kb. I would like to try to use RACE to >amplify the 5' of this message. There are some kits available that I have >seen advertised. I would
just like to know if somebody has ever have any >experience using these
kits and which kit is best based on your experience.

> Thanks in advance.

I have used with success the BRL 5' RACE kit.  My only gripe is that
the anchore primer they include contains uracil (part of a UDG-based
PCR cloning system of theirs) which means, I think, that for optimum
efficiency during the transformation step you really should have the
complementary uracil adapter on your gene-specific primer, followed by
UDG treatment and cloning into BRL's specific vector.  I didn't do all
that and it still worked albeit at a rather poor efficiency.

John



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