3' RACE - looking for suggestions

[Ron Flannagan]

I'm using the GibcoBRL 3' RACE kit and having no luck in amplifying the
3'end of a gene.  With the information I have, the 3'end is 2-3kb away from
my upstream primer.  Is that too long of a sequence to try to use 3'RACE
for?  I am using Superscript II RT and have tried many different cycling
parameters all with no luck.  I have also adjusted the Mg concentration.
Anyone have any experience and suggestions that would help?  Thanks in
advance.


Ron Flannagan
rflannag at magnus.acs.ohio-state.edu