I have been trying to clone my gene of interest using "rapid
amplification of cDNA ends (RACE)". I purchased the cDNA
(double-stranded) from Clontech. So I did my first set of PCR using
adaptor primer and my gene specific primer to perform 5' RACE or 3'
RACE. But I got no bands. When I perform a second set of PCR (nested
PCR), I got basically smears on the gel. I varied temperatures and
cycling programs etc., but I still got smears instead of distinct
bands. So I would like to ask if anyone has done similar things or if
anyone has some suggestions.
Any suggestions will be appreciated.