Techne PrimeQ - Manual (Page 83)

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2. Passive reference dye (PRD) correction:
• Check the box if this correction method is required (only available if a PRD was selected in
the dye usage table). The purpose of the PRD is to normalize the reporter fluorescence and
make well-to-well comparisons more accurate. The readings are normalized by dividing the
fluorescence of the reporter in each well by that of the PRD. This correction method is
applicable to all the analysis types available on PrimeQ (see section 4.4 for more details).
• Click Next.
3. Baseline correction method:
• This contains a number of options and allows the user to adjust the data for any background
fluorescence. The correction uses the fluorescence readings in the early cycles of the PCR
while fluorescence levels are low, averages out the early noise and subtracts it from
subsequent readings. As with PRD correction, baseline correction can be useful for
correcting the data prior to performing an analysis and so helping to increase the accuracy
of the assay. See Chapter 4 for more details.
• Click Next.
4. Set the analysis parameters:
• Select suitable parameters for the analysis type chosen e.g. Cq calculation method, which
reads to use for end-point scoring etc. This screen helps the user decide how to choose the
parameters and/or set thresholds accordingly.
5. Report options:
• Clicking Next leads through to a window that provides options for choosing which graphs
and tables appear in the report. Separate analysis methods and their report options are
discussed in Chapter 4. The user can choose to select the option to run straight through to
report – see section 3.7.6.3 for more information.
6. Summary:
• Clicking Next takes the user through to a page summarizing the settings. Change any
details using the Back function.
• Click Finish to complete the setup and return to the Experiment Editor.
Once the Analysis Wizard has completed, the Experiment Editor pane is updated with the selected
analysis method.