To demonstrate that you are confortable with the molecular and biochemical concepts associated with the PCR reaction, answer the questions below. Your answers are due on Canvas. Show your work and explain your logic in complete sentences. Please note that the deadline for this assignment is indicated on Canvas as well.

A 50 μl PCR reaction contains two oligonucleotide primers. Each primer is present in the PCR reaction at an initial concentration of 1μM.

a) How many moles of each primer are present in the PCR reaction?

There are 5.0x10^-11 moles of each primer present in the PCR reaction. A concentration of 1μM suggests that the solution contains 1 micromole (µmol) of solute in 1 liter of solution. In the PCR reaction, the moles of each primer multiplied with the total volume of the PCR reaction can give the total moles of each primer.

b) How many molecules of each primer are present in the PCR reaction?

There are 3.011x10^13 molecules of each primer present in the PCR reaction. There are 6.022x10^23 molecules in 1 mole, so, multiplying moles times Avogadro’s constant gives the answer

c) If the size of the PCR product in the above reaction is 500 bp, and the PCR reaction yields 2μg of the product, how many molecules of each primer are consumed by the PCR reaction?

HINT: Assume the average molecular weight of a nucleotide base pair is 650 Daltons.

d) What ratio (percentage) of each primer was consumed by the PCR reaction?

e) Another primer set amplifies a 200-bp fragment. Assuming that the ratio of primers used remain the same for reaction using this new set of primers, what would be the yield of the reaction? (i.e. what mass of 200-bp product is generated?)