Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 21)

Section 2 Quick Guides
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3. Select the wells to analyze by clicking Analyze Wells.
4. Define the positive and/or negative controls in the Define Controls column within the End
Point Analysis table.
5. Click Recalculate. The End Point Analysis table displays a positive, negative, or blank label
for each unknown under the Unknowns Call column.
6. Click Reports to obtain customized reports of the End Point Analysis.
End Point Analysis of an Existing Data File
1. Within the Workshop module click Data and select your desired data file using the file
tree browser.
2. Click Analyze.
3. Click the End Point tab.
4. Make selections for the following parameters:
• Method
•
End Point Tolerance and Tolerance Parameter
5. Select the wells to analyze by checking Analyze Wells. Click on Analyze Selected Wells,
and close the Select Wells for Analysis floating window when finished.
6. Define the positive and/or negative controls in the Define Controls column of the end
point analysis table.
7. Click Recalculate. The End Point Analysis table displays a positive, negative, or blank
label for each unknown under the Unknowns Call column.
8. Click Reports to obtain customized reports of the End Point Analysis.
2.4.3 Allelic Discrimination Quick Guide – For Multiplex Data Only
The Allelic Discrimination feature of the Data Analysis module is available post-run and offers
flexibility for analyzing allelic discrimination data from multiplex PCR experiments. You can display
samples on a scatter plot based on threshold cycle or relative fluorescence units (RFU) values at
any PCR cycle. You can have the iQ5 software automatically make allele calls, or you may
manually make the allele calls.
To analyze an allelic discrimination file:
1. Within the Workshop module, click Data, and select your desired data file using the file
tree browser
2. Click Analyze.
3. The file opens in the PCR Quant tab of the Data Analysis module. The iQ5 software
automatically chooses the Data Analysis conditions. To manually adjust these conditions,
refer to the PCR Quant Tab.
4. You can select or deselect wells that will be included in the analysis by selecting the
Analyze Wells checkbox.
5. Click the Allelic Disc tab in the Data Analysis module.
6. Select the Fluorophores that represent Allele1 and Allele2 in the Assign Fluorophores
area.