Bio-Rad Firmware & Software Updates - Manual (Page 111)

Data Analysis Windows
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Adjusting the End Point Data Analysis
Adjust the information shown in the End Point tab by using these methods:
• Choose a Fluorophore from the pull-down list to view the data
• Choose an End Cycle to Average value to set the number of cycles that the
software uses to calculate the average end point RFU
• Select RFUs to view the data in relative fluorescence units
• Select Percentage of Range to view the data as a percentage of the RFU range
• Select wells in the well selector to focus on subsets of the data
• Select a well group (page 75) to view and analyze a subset of the wells in the plate.
Select each well group by name in the Well Group pull-down menu in the toolbar
Data Description for End Point Analysis
Table 32 list the types of information shown in the spreadsheet in the End Point tab.
Allelic Discrimination Tab
The Allelic Discrimination tab assigns the genotypes to wells with unknown samples. Use this
data to identify samples with different genotypes, including Allele 1, Allele 2, Heterozygote, No
Call (no amplification), or Undetermined.
NOTE: The data for allelic discrimination must come from multiplex runs with at
least two fluorophores. Each fluorophore identifies one allele in all samples.
Allelic discrimination analysis requires the following minimal well contents:
• Two fluorophores in each well
• NTC (no template control) samples for optimized data analysis
The software displays allelic discrimination data in these layouts:
• RFU chart. View the data in a graph of RFU for Allele 1/Allele 2. Each point in the graph
represents data from a both fluorophores in one well. You may toggle between Cartesian
or Polar coordinates by checking and unchecking the Polar Coordinates check box.
Cartesian Coordinates represents RFU for Allele 1 on the x-axis and RFU for Allele 2 on
the y-axis. Polar Coordinates represents the angle on the x-axis and the RFU distance on
the y-axis from the origin (median of all NTC).
• Well spreadsheet. Shows a spreadsheet listing the allelic discrimination data collected
in each well of the plate
• Well selector. Select the wells with the end point data you want to show
Table 32. End Point spreadsheet contents.
Information Description
Well Well position in the plate
Fluor Fluorophore detected
Content A combination of the Sample type and Replicate #
End RFU RFU at the end point cycle
Call Positive or Negative, where positive samples have an RFU
value greater than the average RFU of the negative controls
plus the Cut Off Value
Sample Sample Name loaded in the Plate Editor