| | Re: how to dilute 100mM dNTP to 1.25mM? what is the calculation to make pcr reaction.
The best and the most used formula I ever learned at school is this one:
c1= the concentration of the solution you want to dilute
v2= the volume you should use of the solution
c2= the concentration you want in the final solution
v2= The volume you want of the final solution
So lets say you want to make 500 ul of a 12,5 mM solution the formula is:
100 mM * v1 = 12,5 mM * 500 ul
v1 = 62,5 ul
That means you have to use 62,5 ul of your 100 mM solution and dilute it in 437,5 ul of whatever you want to dilute it in to get a final volume of 500 ul. Then you have a 12,5 mM solution of your primers, and you can then use 2,5 ul of this solution in your 25 ul to get a final concentration in the reaction of 1,25 mM.