Question

When you order primers, they are delivered as dried (lyophilized) DNA in an Eppendorf tube. You must then dilute these primers with molecular biology grade water to an appropriate concentration. Typically, the primer tube simply lists the mass of DNA that is in each tube. Based on this information, complete the following table to create 100 uM stock primers for the 6 primers

Primer	Number of nucleotides	Mass (ug)	nMoles	H20 needed to create 100 uM stock
Polymerase Forward	20	206
Polymerase reverse	20	174
Sybr Forward	20	167
Sybr Reverse	20	231
TaqMan Forward	20	100
TaqMan Reverse	20	149

Answer 1

Answer- The nmol of DNA can be determined by the formula ugDNA× pmol/660pg × 10^6pg/pmol × 1/N and then total divided by 1000

N- number of nucleotides

660pg/pmol- average weight of nucleotide.

Primer	Nucleotide	ug	nmol
Polymerase F	20	206	15.60
Polymerase R	20	174	13.18
Sybr F	20	167	12.65
Sybr R	20	231	17.50
Taqman F	20	100	7.50
Taqman R	20	149	11.28

The dilution is done by multiplying the nmol of DNA by 10 and adding that amount of water.

So the amount of water that will be add to different primers are respectively from upside to down - 156, 131, 126, 175, 75, 112 ul of labgrade water.