Can a wrong input of RNA and standard curve in a qPCR, resulting in lower RNA concentrations in the mix, be corrected afterwards?

04 December 2020 1 9K Report

I performed a qPCR in which I had an input of 5uL RNA as well as the standard curve instead of the 10uL that was supposed to be the input. The mix (15uL) that was made was based on a 10uL input. Is it possible to perform corrections afterwards for the lower RNA concentrations?

Jochen Wilhelm

Simply use half of the planned concentration (i.e. the actually used concentration) for the calculations.

If you prepared the standard curve not for quantification but only to estimate the amplification efficiency, the actual concentrations are not relevant. Only the dilution factors are relevant, and those didn't change.

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