RACE to get full lengh cDNA sequecne for gene with trancsript variants?

06 June 2019 1 1K Report

Hi, I have six transcript variants for my gene of interest for which I am doing RACE (both 5'RACE and 3'RACE). I have a problem in understanding that when I clone, for ex. 10 clones from each 5'-RACE and 3'-RACE and then I do sequence alignment that will give me the idea for different transcript variants expressed in cell but how would I combine the 5'-RACE and 3'-RACE sequences to get full length sequence for each transcript variants that expressed in cell?

Cecil Saunders

You should be able to use the reverse compliment of primers you designed for the 5' and 3' Race to do a gene specific PCR. Once you have sequenced that amplicon it should just be a matter of matching up the part of the sequences that correspond to the primers.

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