A student needs to add 2.2U of Taq polymerase into a reaction that will have a f

Question

A student needs to add 2.2U of Taq polymerase into a reaction that will have a final volume of 25ul. The Taq is stored at a concentration of 5U/ul. How much Taq Polymerase should be pipetted?

4.4ul

.44ul

10ul

A student is preparing to set up a 15ul PCR reaction and is trying to calculate how much primer needs to be pipetted into the reaction. The student is using a reagent that has both the forward and reverse primer together and the concentration of each primer in the reagent tube is 15uM. The protocol says to “Add forward primer to a final concentration of 1.5uM. Add reverse primer to a final concentration of 1.5uM”. Knowing that the primers are mixed together in one tube, how much of primer mix should be pipetted in order to have the forward and reverse primer at the correct final concentration.

1.5ul

3.0ul

15ul

Explanation / Answer

1) 2.2 U needs to be added to 25microlitres

so, per microlitres will be = 2.2/25=0.088U

in concentration, 5U/ microlitre= 0.088*5= 0.44microlitres

2) for 15ul reaction, 1.5uM needs to taken

c1v1=c2v2

15uM v1= 1.5uM* 15microlitres = 1.5mirolites

since reverse and forward primer are mixed together, collectively, 3.0microlitres

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