Hi
I'm doing my master thesis and having a hard time optimizing my PCR reaction. I'm trying to add 2 concentrations of MgCl to my mastermix. I already have 1,5 mM MgCl in my master mix but need it to be 2 mM and 2.5 mM respectively, before I find another hotstart, but can't for the life of me remember how to calculate in moles etc.
I have an end concentration of 25 µL in my wells.
I already have 1.5 mM MgCl
My stock has 50 mM MgCl.
How many µL of MgCl am I supposed to add for the two PCR's?
Would like to understand the calculations.
Can someone help?
Thanks in advance
You can use the equation v1C1 = v2C2 but you already have 1.5mM Mg in your master mix so just use the difference in concentration in the equation. So to get 2mM in a solution that has 1.5mM we just need an increase of 0.5 mM so ci is 50mM and c2 is 0.5mM and V2 is 25ul per pcr rection so the additional volume V1 is c2v2/c1 so the additional volume of stock Mg for 2 mM ( 0.5mM extra) is 0.5x25/50 or .25ul stock per pcr reaction
- Badges
- Science method