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Volume of DNA to add to PCR Master Mix? - (Sep/05/2009 )

Hi all,

I have figured out the concentraton of DNA in each of my samples. I want to have equal amounts (1 microgram) of DNA in each of my samples. How do I calculate the amount of DNA template/water to add to the master mix? I want 30 microliters of PCR product (I already know how much primers, taq, etc. to add).



Hi EbolaZaire -- welcome to the BioForums!

If you know the concentration of DNA in each of your samples, then you have a measure of the mass of DNA per volume of liquid. What volume of DNA solution at that concentration will contain 1 microgram of DNA? That is the amount you need to add to your reactions, and bring them to a final volume of 30 microliters with sterile water.