Sequencing 3'RACE PCR products: help on dTVN primer?
Thu Aug 14 16:51:45 EST 1997
Having a nice summer everyone?
My group is doing high-throughput (400-800 PCR products per day)
sequencing of 3'RACE PCR products with TaqFS and dye terminators. The
products are easily sequenced from the 5' end with a standard primer. I
want to obtain sequence from the 3' end as well. The RT-PCR products
have very long stretches of polyAs that cannot be read through using the
3' end primer. Can one use an anchored polyT primer such as dTVN, where
V= A,G, or C? I have tried this, but have had no success - multiple
priming, just plain messy data. Does anyone have any suggestions as to
what can be done?? Reaction conditions, etc.???
Sincerely Stuck at the 3' End,
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