The "RACE" for 5'ends

[ajwatson]

In article <2mc8h0$pfj at mserv1.dl.ac.uk>
"Matthias Zeiner" <mzei at sun0.urz.uni-heidelberg.de> writes:

> Did anybody have success with Clontech's amplifinder kit?
> I know there is a similar kit from BRL, is that one ok?
> How could I improve the "RACE" technology?

I have used the GIBCO/BRL 5' RACE kit with good success to clone the 5'
end of a G protein beta subunit cDNA.  My only complaint about the kit
is that the anchor primer supplied contians uracil residues for
(downstream) cloning using a uracil deglycosylase system sold by (guess
who) GIBCO/BRL.  Transformation of uracil-contianing plasmids into dut+
strains leads to rearrangements, etc.  I found a reduced number of
clones upon transformation, but was able to fish out the few that had
my insert.  Interestingly, when I made an anchor primer that was
lacking the uracil stuff, it didn't work at all!

John Watson
Caltech Biology
ajwatson at romeo.caltech.edu