Question

PLEASE HELP WITH CHEMISTRY PROBLEM

you need to treat your PCR product with Fnu4HI (final concentration = 0.1 U/µl) in NEB buffer 4 (final concentration = 1X). For a total reaction volume of 30µl, calculate the volumes of Fnu4HI stock and NEB buffer 4 stock that are needed.

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Info about stocks:
Fnu4HI stock solution is 1 U/µl
NEB buffer 4 stock solution is 10X

Answer 1

We have 1U/ul and we need 0.1 U/ul for volume of 30 ul.

we use dilution formula M1V1 = M2V2 , 1 x V1 = 0.1 x 30

V1 = 3 ul ,

now for NEB , 10X x V1 = 4X x 30

V1 = 12 ul

Thus 3 ul of Fnu4HI stock solution and 12 ul of NEB buffer stock solution taken and then mixed and then diluted to 30 ml mark by water. Thus we get required solution.