primer mix

[molbio123]

I want F and R primers to be at 20uM individually in my primer mix.

If I mix 20uM F with 20uM R in equal volumes (1:1), this will make 10uM F/R primer mix, but the individual primers are still at 20uM in the mix? Is this correct? Thanks.

[mmorgan]

Originally Posted by molbio123

I want F and R primers to be at 20uM individually in my primer mix.

If I mix 20uM F with 20uM R in equal volumes (1:1), this will make 10uM F/R primer mix, but the individual primers are still at 20uM in the mix? Is this correct? Thanks.

No, the individual primers are 10uM in the 1:1 mix. You've diluted each primer concentration in half.

For 20uM of each you need 40uM of both F and R and then you mix them together 1:1 for a 20uM concentration of each.