Increasing the dNTPs in a Multiplex Rxn - (Sep/09/2008 )

If I were running a QPCR reaction with only one primer and probe set and I decided to add a second set of primers and probe, by how much would I increase my dNTPs. ie: double them?

Thanks

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why not try use master mix for qPCR?

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Do NOT add excess dNTPs, unless you add an equilibrate Mg2+, as each of the dNTPs chelate Mg2+ at the ration of 1:1 (eg. 5 mM dATP chelate 5 mM Mg2+, and 5 mM dNTPs totally chelate 20 mM Mg2+). It is well known that Mg2+ is critical to PCR, so reduced concentration of free Mg2+ may dramatically decreased the efficiency of PCR. By the way, 0.2 mM dNTPs final, the most usually used concentration of dNTPs, is sufficient for almost all kind of PCR.

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