long PCR with dUTP

Timo Hiltunen bltihi at uta.fi
Tue Feb 25 01:56:38 EST 1997


Michael C. Gorry <mcgorry at med.pitt.edu> wrote:
: 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?  
Hello!
Although I have done only shortish PCRs (< 1 kb) with dUTP,
it is my experience that dTTP gives more sensitivity and productivity.
Thus, in ordinary and easy PCRs, I use dUTP but switch to dTTP if
more "power" is needed.
(I have not tried to raise dUTP concentration above that of other dNTPs.)
TH



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