PCR [dNTP] - dNTP concentrations (Mar/19/2009 )

This is the thing (I know this must sound very simple, but I would honestly appreciate some help).

I take 5 ul of dNTPs (1mM stock solution) to put into a PCR reaction --> I suppose this is 0.25 mM of each dATP, dCTP, dGTP and dTTP (¿?)

PCR reaction = 50 ul

According to my calculations (C1V1=C2V2) --> C2 = <(1000 pmol/ul)(5 ul)> / 50 ul = 100 pmol/ul
That is, the concentration of dNTPs in the 50 ul PCR reaction is 100 uM. This in turn means that I have 25 uM of each individual dNTP (i.e., dATP, dCTP........dTTP) in the 50 ul PCR reaction.

Is all this correct or am I completely ¿?

I am comparing two protocols, and 25 uM of each dNTP looks a bit low.

THANKS EVERYBODY FOR YOUR HELP !!!!!!!!!!!!!! GOD BLESS YOU ALL !!!!!!

-autoclave-

That does seem pretty low. I use the Qiagen taq mastermix and I think that's 200 uM per dntp.

-microgirl-

I will say the 1mM stock is 1mM of each dNTP, not 1/4th each dNTP, however you should double check this.