PCR with Poly A primer

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

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.

More information about the Methods mailing list