I have a question about the quantification through qPCR. The my question is: if I made the qPCR of a fungal functional gene, how is possible to obtain from the quantification number (in nanogram) the equivalent number in number of nuclei?
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$\begingroup$ To clarify: you're asking about converting between copy number (which may be expressed in moles, molarity, or just a raw number) and nanograms of DNA? $\endgroup$– PunintendedFeb 4, 2020 at 23:44
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$\begingroup$ If I obtain the quantification between nanogram to covert this number in number of nuclei. Because in one paper I found this sentece: "As few as 148 fg of Verticillium dahliae DNA were detected and quantified (through qPCR), which is equivalent to five nuclei". How is possible to make this convertion? @Punintended $\endgroup$– GiorgiaFeb 5, 2020 at 8:44
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From your comment:
"As few as 148 fg of Verticillium dahliae DNA were detected and quantified (through qPCR), which is equivalent to five nuclei". How is possible to make this convertion?
The Verticillium dahliae JR2 genome is 36.2 Mb. Assuming this organism is haploid and dsDNA has an average mass of 660 g/mol per base pair, that means the molar mass of the genome is 2.40 × 1010 g/mol. Therefore, 5 nuclei will contain 8.30 × 10-24 mol of your genome. Multiplying the molar mass by the number of moles gives us a mass of 1.99 × 10-13 g, or 199 fg, which is close to your cited value.
