Prokaryotic cDNA Cloning

Tue Aug 31 15:25:12 EST 1993

We are working on cloning several prokaryotic genes whose expression we can
induce.  I was wondering if anyone has experience in taking prokaryotic RNA,
tailing it with poly(A) polymerase, making cDNA, and cloning it.  I assume this
 is possible.  We already know some N-terminal DNA sequence for a couple of
these and I believe that using poly(A) tailing and RT-PCR with poly(dT) and an
upstream primer, we could get the C-terminal end of these genes.  Any
suggestions?  Conditions?  Best enzymes?  Can it be done?

  Tom MacAllister
  n490047 at UNIVSCVM

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