Hi I am a bit confused.
They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8 ng/uL. Then I need to calculate the volume of water required to bring the total reaction volume to 50 uL.
They gave the different volumes for the PCR reagents which was; (PCR master mix-25, Forward primer -0.5, reverse primer - 0.5) Now i need to figure out the DNA volume and the water volume so that the total equals to 50 uL. But how am I to figure out the volume
Just to add to the excellent answer, in order to prepare the 1:10 dilution you must put 1ul of your DNA sample and 9ul of water.
According to the quantity you obtained, you can prepare 100 ul dilution with 7.5ul DNA(+92.5ul water). Then you can pick 1ul from it (30ul DNA) or 2ul DNA for the reaction. Finally, you can add 23 ul water (for 1ul picking) or 22ul water (for 2 ul picking), making your final volume 50ul (25 MM+0.5 P1+0.5 P2+ 2 dil. DNA+ 22 water).
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