Researchers performing qPCR will often create a standard curve based on nanograms of amplicon, and then need to convert the resulting nanograms detected to copy number. The formula for making this conversion is as follows:
Where:
X = amount of amplicon (ng)
N = length of dsDNA amplicon
660 g/mole = average mass of 1 bp dsDNA
† The actual oligonucleotide MW (in Daltons or g/mole) is provided on the IDT Spec Sheet for each oligonucleotide and can be substituted for this arithmetic phrase, which only provides an average MW for a sequence of this length.
An alternative is to create the standard curve based on copy number; e.g., ranging from 102 to 107 copies, in 10-fold increments. You can then determine copy number directly from the standard curve.
Let a free copy number calculator do it for you!
A convenient, free-access copy number calculator to convert nanogams to copy number for real time PCR (RT-PCR) was developed by former IDT customer, Andrew Staroscik, who has since left the bench.
The copy number calculator can be found at: sciprim.com/html/copyNumb.v2.0.html.