long PCR with dUTP

Michael C. Gorry mcgorry at med.pitt.edu
Mon Feb 24 18:20:36 EST 1997


I am trying to amplify a longer DNA (4kb) fragment using dUTPs instead
of dNTPs.  I have been unable to get an amplification product with U but
have obtained it with T. Is there a limit on the amount of U that can be
incorporated?  I have not been able to find anything out about it and before
putting more effort into it, I would like to know if anyone has any information.
To date, I have tried taq, klentaq, pfu, and rTth.  I had expected the best
results with the rTth, but was unable to reach the 0.6kb control with any of the
polymerases used.

Thanks for any help.

Mike

-- 
Michael Gorry
mcgorry at med.pitt.edu



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