MadSci Network: Molecular Biology |
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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