I'm doing absolute quantification for my qPCR for virus infection and ran my samples alongside a standard curve as well as GAPDH for a reference gene. I understand how to get copy number of my target gene from the standard curve but how to I (or do I?) incorporate my reference gene into my calculations?
Hi,
I doubt that this approach will help you because you are looking for the virus copy number if I am not mistaking. GAPDH will rather work as a loading control because your infeciton might also affect the ref genes. You could use a calibrator to normalize the viral load to the input amounts which could be an internal control or so.
Svne
Hi Molly Braun
I agree with Jack Greenhouse . Just divide the results of absolute quantification by GAPDH copy number. You have a DNA or RNA virus?
Best,
Jacó
I am not familiar with GAPGH. But I know you can make standard serial dilutions from your gene of interest then generate a standard curve. That is it.
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