5'-RACE

Shiao Wang Shiao.Wang at usm.edu
Thu Nov 19 02:05:51 EST 1998


If contaminating dsDNA was tailed at the 3'end with C, both strands
would serve as template when (GGG)n is used as primer. However, the
newly synthesized strand (containing GGG) would not be a useful template
for further PCR unless it contains an annealing site for the gene
specific primer. Thus, am I correct in thinking that if the the
contaminating DNA lacked annealing sites for the gene specific primer,
the DNA would not be a problem?

I know several students who have had success with 3'RACE but are
struggling with 5'RACE. Other than the fact that longer cDNAs are more
difficult to obtain than short cDNAs, is there a good reason for this?

Recommendations of references on 5'RACE that you think were particularly
helpful would be appreciated also. Thank you for your attention.

-- 
Shiao Y. Wang
Department of Biological Sciences
University of Southern Mississippi



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