I'm a little confused, I am doing a PCR reaction that requires 100ng of genomic DNA template, and the concentration of my genomic DNA is 591,9ng/ul. So is 100ng of the template the same as saying 100ng/ul? In that case, I would use C1V1=C2V2 to work out the volume of genomic DNA to use. Is this correct? I am using a 20ul reaction.
Or should I use v=m/c, so the answer is 0.167ul?
100ng is an absolute amount but 100ng/ul is a concentration and the 2 are different. Your dna ia 592ng/ul so 1 ng is 1/592 ul of this so;ution. Your pcr needs an absolute amount (weight) of dna of 100ng so you need (1x100)/592 ul of dna sample which is about 0.17ul. This is too low a volume to pipette so I would dilute 1ul of dna to 10ul of solution and pipette 1.7ul into the pcr mix
Hi Danijela Hw
I agree with Paul Rutland , only that while diluting your stock DNA, do 2uL in 18uL (1:10 dilution) and take out 1.7uL for PCR.
Pipetting 2uL out of stock rules out pipetting errors to some extent.
Subham.
Thank you so much Paul Rutland and Subham Basak for your answer! It helped me a lot!
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