PCR with Poly A primer

jon boyson boyson at primate.wisc.edu
Wed Sep 27 13:21:54 EST 1995


Hi--
We use this procedure with good results. The only difference is that I 
think your poly A primer will not give you good PCR results (so I've 
heard). If it's not too much trouble make the following modifications to 
your poly T RT primer:

1)Make the last base G, C, or A (ie. mixed base) to ensure your primer 
primes from the same place (beginning of poly A tail) all the time.

2)Stick an irrelevant oligo on the 5' end of your poly T primer (w or 
w/o restriction sites etc) so that when you come back to amplify the 
cDNA you will use this primer instead of the poly A.

Hope it helps.





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