dNTP problem

Andrei Popescu andreip at byte.cas.vanderbilt.edu
Mon Aug 28 17:16:45 EST 2000


Hello everybody,

I have been trying to get a PCR started for 1 month, no results. I
modified everything, especially annealing temperatures and DNA
concentration. I took the template and primers to another lab, and boom,
the reaction worked in 4 out of 4 samples. I came back and repeated in
my lab but using dNTP mixture from another lab, and it worked
beautifully. I tried in parallel with my old dNTP-nothing, not even the
positive controls. I bought new dNTP from promega, 4 vials - nothing
again. I used same .25 mM concentration each dNTP, in successfull or
unsuccessfull reactions. So it seems to depend on whether the dNTPs are
premixed or mixed by me. Either I am doing something systematically
wrong when mixing , or there is something more subtle. At 200 $, I
cannot buy dNTPs every day. Has anyone found something similar?

Thanks in advance!

Andrei Popescu, RA
Molecular Biology
Vanderbilt University


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