In case of no passive reference dye(ROX) in a qpcr experiment, how instrument handles background correction? We are using ThermoFisher's QuantStudio 3 and 5 instruments.
If instrument does not do any background correction in this case, how the data should be processed for further analysis?
Normalizing the fluorescence intensity is accomplished dividing the emission intensity of the reporter dye by the emission intensity of a reference dye. Without background correction using ROX fluorescence, data looks off the charts.