Hi all,
is this following calculation correct ?
I need 0.5 ug of DNA for a restriction digestion and I have an initial concentration of DNA of 696 ng/ul. The total volume of the restriction digestion reaction has to be is 25 ul.
So I calculated first the concentration of DNA corresponding to 0.5 ug
C= 0.5 ug/25ul = 0.02M
and then the initial volume to take from the DNA extract to add to the digestion reaction .
Vi= 0.02M x 25 ul/ 0.69 M= 0.7ul
Thanks
you need 500ng and the concentration is 696 ng/ul so you need 500/696 ul which is 0.718 ul of stock dna.. The reaction volume is not important when you are dealing in weight only when you are working in units of concentration.
Dear Silvia,
If you are doing a single digestion:
DNA = 0.72 ul
Restriction Enzyme = 1.00 ul
10X Buffer = 2.50 ul
Water = 20.78 ul
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