How do you calculate how many moles of product you have in your total pcr reaction. Then how do you calculate how many molecules of your target are in the total reaction volume? Then I need to be able to calculate the average target-amplification factor per cycle of PCR. Any help would be greatly appreciated. Thanks

Okay
Let’s do some math
1. what is the mol wt of PCR product
Say you have a 100bp PCR product
1bp= 660 Da
100bp= 66000 Da

by definition one mole of PCR product will weigh 66000g and will have 6.023* 1023 molecules (Avogadro’s number)
A one molar solution will have 66000g of PCR product in a Litre

2. amount of PCR product (measure using a spectrophotometer)
you will have the value in x nanogram/microlitre
let us say it is 50nanogram/microlitre

i.e 50nanog/microL
or 50micro/mL
or 50millig/L
or 50*10-3g/L


3. Now we know that
66000g/ L is 1 molar solution
1g/L 1/66000 molar
50*10-3g/L 50*10-3/66000 molar= 750 * 10-9 molar or 750 nanomolar

4. The other way of doing of calculating moles would be:
Say you have 100 microliter of PCR product at 50nanogram/microlitre
So you have 5 microgram of your product

Total mass/ molar mass= number of moles
5* 10-6 g/66000=75* 10-12 moles
75 picomoles of PCR product

5. 1 mole =6.023* 1023 molecules
(10 raised to 23)
75* 10-12 moles = 6.023* 1023 *75* 10-12
= 4.51 *1013 molecules