I would assume the "400 uL primer master mix" is a 400 uL master mix

then the final volume is 406

so the final concentration of the primer would be 0.985... and 1.4777... uM

usually when we do PCR, we will aliquot and dilute the primer stock (the one with 100uM concentration) down to 10 or 5uM or as needed then we use it.

Sophie Gonzalez,

As suggested by my colleague Jonathan Jonathan, I think that you have first to prepare a primer working solution from the mother solution (100µM in your case) at the protocol preconized final concentrations. For example, a 100µl volume of working solution for both primers at 10µM could be prepared by adding 10µl of 100µM to 90µl of water.

In your case, volumes of 4µl and 6µl of 10µM working solution will give you respectively final concentrations of 100nM and 150nM in a final volume of 400µl.

Jonathan Jonathan

I need to know concentration of individual primers. The total primer mix volume would be 400uL, that’s including water and primers.

Aissa Saidi

I’m not sure about bringing the primer concentration to 100uM to 10uM. I just know as of right now when pipetting directly 4uL or 6uL, it would go into a total volume of 400uL. There will be many other primers, but right now I need concentrations of individual primers and I can’t tell if my math is right.