Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 84)

Section 6 Data Analysis Module
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Reference: Livak JL, Marmaro J and Todd JA, Towards fully automated genome-wide
polymorphism screening,
Nature Genetics , 9, 341-342 (1995)
6.7.10 Restore Default
At any time, all modifications to the allelic discrimination data can be reversed by clicking
Restore Default. The iQ5 software will reload the original x-axis allelic 1 fluorophore and y-axis
allelic 2 fluorophore data, original display mode and Show Labels setting that were originally
saved in the .opd file and will also set the Automatic Call method to recalculate the data, plot the
chart, and display the results in the allelic discrimination data spreadsheet.
6.8 End Point Analysis
The End Point analysis module is a convenient method of analyzing final relative fluorescence
unit (RFU) values.
End point analysis can be performed in two ways:
•
Selecting the End Point tab in the Data Analysis module for an existing data file
•
Click Run End Point to initiate the collection of End Point data from a sample plate
Newly collected End Point data is displayed immediately following an End Point run. Any existing
.opd data file can also be viewed in the end point module however, the corresponding PCR
quantification data must be analyzed in either the PCR Base Line Subtracted Mode or the PCR
Base Line Subtracted Curve Fit mode before the End Point tab becomes available. Although any
file with amplification data may be analyzed post-run in both the PCR Quant and End Point tabs,
an End Point run may only be analyzed in the End Point tab. Melt Curve-only experiments, which
neither contain amplification data nor End Point data, may not be analyzed in the End Point tab.
6.8.1 The End Point Analysis Settings
The End Point tab is comprised of several sections, each one described in detail below. The
Display Wells and Analyze Wells buttons have the same functions as described in section 6.2.4.
Method Box: End Point Analysis Methods
Below the file and fluorophore information is the Method box. The Method box allows you to
select the method of assigning positive and negative values to your unknowns based on RFU
values. The Method box consists of the following three choices:
•
Negatives: This is the default method. Select this method to use negative controls to
define or call unknown samples. Samples are considered positive if they are greater in
RFU value than the negative control average plus the tolerance
•
Positives: Select this method to use positive controls to define or call unknown samples.
Samples are considered positive if they are greater in RFU value than the positive control
average minus the tolerance