RACE troubleshooting

Corinne RENIER crenier at leland.stanford.edu
Mon Feb 11 15:15:42 EST 2002


Hi,

Does anybody have experienced problems with the SMART RACE cDNA
amplification kit (Clontech)?
After the first strand synthesis, the RACE PCR using my gene specific
primer (either 5' or 3') failed to give me clear band (I get smears),
while my positive control (using 5'+3' gene specific primers) on the
cDNA RACE populations (either 3' or 5' RACE cDNA population) is working
under this PCR conditions (I've got a clear and nice band).
I've designed different primers, but each time it's the same story, so I
guess the primers are ok! I've been following the troubleshooting guide
advices without success!
Any advices or tricks to suggest? Does anybody have experienced with
other RACE kits to share?
Thanks

Corinne




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