Paul.Cullen at med.monash.edu.au
Mon Aug 28 20:57:35 EST 2000
Make sure you are not defrosting them in a water bath. dNTPs are quite
sensitive to heat and should be stored at -20 C. Defrost on ice.
Andrei Popescu wrote:
> 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
Bacterial Pathogenesis Research Group
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