MadSci Network: Molecular Biology

Subject: How to calculate the theoretical yield in moles in a PCR?

Date: Thu Feb 22 16:19:27 2007
Posted by Kanwaljit S Aulakh
Grade level: undergrad School: No school entered.
City: No city entered. State/Province: No state entered. Country: No country entered.
Area of science: Molecular Biology
ID: 1172186367.Mb

For a particular PCR amplification, suppose that the initial concentrations of 
both For and Rev primers were 1.20 micromolar, each dNTP was present at a 
final concentration of 100 micromolar, Mg 2+ was included at 2.5 millimolar 
along with 0.10 ng of a 3,000 bp template in a final volume of 100 
microliters. What is the limiting reagent? What is the theoretical yield (in 
moles) of an 600 bp PCR product under these conditions (assuming equal 
quantities of A, C, G, and T were incorporated into the PCR product)? If a 
base pair has an average molecular weight of 650 g/mole, how many micrograms 
could be produced? Be sure to show your calculations. Note that comparing this 
number to your yield provides an estimate of the efficiency of your PCR 
amplification. 30 cycles of PCR are performed

Basically I tried everything but what weight do I use for finding out the 
moles of the 600bp? Do I use the 0.1ng? I came up with 2.57e-16 moles and 
107621.14 micrograms

Re: How to calculate the theoretical yield in moles in a PCR?

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