Rotor-Gene Q MDx CE User Manual 02/2022
108
with the calibrator sample is calculated based on the takeoff point and the reaction efficiency. This
is expressed in scientific notation.
Note
: The value displayed in
Average Amplification
to the right of the ± represents the standard
deviation of the average amplification, after removal of outlier amplification values. If this value is
large, then there may be a large error in the overall calculated concentration values.
Relative concentrations are calculated by the software as follows:
1.
The takeoff point of each sample is calculated by looking at the second derivative peaks.
2.
The average increase in raw data 4 cycles after the takeoff is calculated. This is the
amplification value for the sample.
3.
Outlier amplifications are removed to account for noise in background fluorescence.
4.
The remaining amplifications are averaged. This is the average amplification.
5.
The average takeoff point is calculated for each calibrator replicate.
6.
The relative concentration for a sample is calculated as Amplification^(Calibrator takeoff –
Sample takeoff).
7.
The result is displayed in scientific notation in the “Rep. Conc.” column of the
Comparative
Quantitation Results
window.
6.6.7
Allelic discrimination
Allelic discrimination uses real-time kinetic data from 2 or more channels to genotype samples. To
perform this analysis, select
Other
and then
Allelic Discrimination
in the
Analysis
window. When
performing allelic discrimination, it is not sufficient to double-click on one channel to analyze
because this analysis is performed using multiple channels simultaneously. To perform this analysis,
either hold down CTRL and click to highlight each channel you wish to analyze, or drag the mouse
pointer over these channels. Once the desired channels have been highlighted, click
Show
. The list
will update to show all the channels on one line, with a checkmark beside them. This indicates that
they will all be used in one analysis. To remove one or more of these channels, right click on the
analysis and select
Remove Analysis...
. Those channels can then be included in another allelic
discrimination analysis. A channel can only be used in one analysis at a time.
Reports
:
This opens the “Allelic Discrimination Analysis” report for preview.
Results
:
This displays the
Allelic Discrimination Results
window. This window is opened
by default when the analysis is first displayed.