Techne PrimeQ - Manual (Page 113)

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3.12 Passive reference dye (PRD) correction
The purpose of the passive reference dye (PRD) is to normalize the reporter fluorescence so that
well-to-well comparisons are more accurate. The readings are normalized by dividing the
fluorescence of the reporter in each well by the PRD for each well. This correction method is
applicable to all the analysis types available on PrimeQ except “None”.
Setup procedure:
For the PRD correction options to be displayed, a PRD must be assigned in the Dye Usage box
accessed from either the Experiment or Results Editor.
If a PRD is assigned, each analysis method will include options for PRD correction.
Clicking on the PAR button of the Raw Data graph brings up the PRD correction options.
The user can choose to display the raw data for the reporter, the PRD or the PRD corrected
reporter.
The user should note that a PRD is not regarded as a reporter. Therefore, the dye chosen as the
PRD will only appear in the Dye to View menu (a function that allows the user to choose which
dye data is being displayed) when None is selected for the Analysis Method.
Raw data for
Reporter and PRD
For each well, calculate
fluorescence of Reporter
fluorescence of PRD
Plot corrected fluorescence
vs. cycle number
Raw data for
Reporter and PRD
For each well, calculate
fluorescence of Reporter
fluorescence of PRD
Plot corrected fluorescence
vs. cycle number