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5' RACE of difficult region

Lirong Zeng zeng.17 at osu.edu
Wed Apr 2 12:50:17 EST 2003

I am trying to get the 5' end sequence of my gene using the Clontech
SMART RACE cDNA amplification kit. It has two very high GC content
(average 82%) region (each ~ 230 bp) at the predicated start coding
region and 5' UTR. I failed so many times. I could  not get RACE
amplification band even when I tried the Clontech Advantage GC PCR kit
and the N7-deaza-dGTP replacement. Do u have any experience on this
kind of problem? Appreciate your information.



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