Fabian.Moebius at uibk.ac.at
Tue Mar 4 13:52:56 EST 1997
We desperately try to isolate the 5prime end of a clone. We already
got 3prime partials. We also cloned the same cDNA from two other
species with some difficulties (most but not all clones isolated from a oligodT
primed cDNA library were incomplete). The problems could be due to a
GC rich 5prime sequence.
We tried RACE with an oligodT primed library from
Clontech). We do not get any 5prime end PCR products whereas 3prime
PCR works. We now tried 5%DMSO which changed the picture with some of our
Seems to be a trivial problem. Nevertheless we fail.
(1) Any suggestions how to overcome GCrich sequences in RACE?
(2) How could we increase the ratio between the desired longer (600bp)
product and an undesired possibly truncated 300 bp amplicon?
(2) Any references for such problem (to make sure that it really is
Thanks for your input!
Dr. Fabian F. MOEBIUS
Institut fuer Biochemische Pharmakologie
Peter Mayr Str. 1
More information about the Methods