PCR with Poly A primer
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.
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