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RACE cloning

RACE cloning - Protocols and Methods Forum

RACE cloning - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 08-07-2003, 07:37 PM
Emily Zhou
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Default RACE cloning



Hi,

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.
Thanks. Emily
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