RACE problem! Please help

ma ddsyr at PUBLIC.EAST.CN.NET
Mon Sep 7 08:41:30 EST 1998


Dear Netters:
I have got a cDNA by RACE method using a general primer (containing
oligoT)as the 3'primer and a degenerate primer as the 5'primer. Now I
want checked this cDNA expression by RT-PCR. But I fail to get the
signal using the same the primers. What wrong? or should I use a
non-degenerate primer (since onw I know the sequence). Should I also
synthesis a new 3'primer, since in RACE, the 3' general primer is behind
the polyA, does that will affect the yield in RT-PCR? Any RACE experts
please urgently give me a help.
I also get another cDNA by above approach, it contains the 5'primer but
no 3'primer. What wrong about that?
Please return to my personnel email.
Thanks
Dr Q. H. Ma
Institute of Botany, Academia Sinica
Beijing 100093, China





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