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You want 50 fmol DNA in a 10 µL reaction. MW of DNA is 607.4 g/bp....

You want 50 fmol DNA in a 10 µL reaction. MW of DNA is 607.4 g/bp. Your DNA is 500 bp in length and at a concentration of 10 ng/µL. How many µL of your DNA should you add to the reaction?

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Answer #1

The information suggests that the size of DNA is 500bp and the molecular weight is 607.4 g/bp. Thus, the weight of available DNA is 607.4*500 or 303700g or 0.31 microgram or 3.1*10-4 ng. The required concentration of DNA is 50fmol i.e. 50*10-15 mol.

Using the formula:

M1V1 = M2V2,

10 * 10-9 * V1= 50*10-15 * 10

Solving for V1, we get V1 =50*10-15 / 10-8

or V1 = 5*10-6 ul or 5 picolitre.

Thus, only 5 picolitre of this stock solution is required to be mixed in a reaction volume of 10 ul to obtain the desired concentration.

Since this small volume cannot be measured using any pipette, serial dilutions of the stock solution are first made until an requisite dilution is reached and this final dilution is used for preparing the reaction mixture.

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