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dNTP problem and magnesium Mg concentration in PCR reaction



dNTP problem and magnesium Mg concentration in PCR reaction

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.

Last update 12-Jan-2006, Rating of 3 votes.

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By MANOJ on 01-Jul-2007
SIR,I WANT TO KNOW THAT CAN WE USE DNA CONCENTRATION IN PCR UPTO 10 ng.IF OUR
RESULTS ARE COMMING VERY WELL.BUT ACCORDING TO THE PROTOCOL DNA SHOULD BE USE IN PCR UPTO 50-100ng.
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By VIKRAM BHATIA on 31-May-2006
i am trying to do pcr optimization for cDNA clone but i failed.i tried more than 10 times but i didntr get.
i am using hot start taq and cDNA clones and M13 universal primer.using annealing temp. 55 for 30sec.
plz suggest me what should i do?
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By E Garcia on 12-Jan-2006
What vendor (brand) sold the dNTP mix that worked? I encountered a similar problem and I found that the Stratagene brand worked better for me.
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