Bio-Rad Firmware & Software Updates - Manual (Page 92)

MiniOpticon Instruction Manual
81
• Select Normalize Data to normalize the RFU data shown in the chart and
spreadsheet
Normalization changes the data on the chart to a range from 0 to 1 on both axes. To normalize
the data, the plate must contain wells with “no template control” (NTC) sample types for both
Allele 1 and Allele 2. For this plot, the RFU data are normalized to the NTC values as a linear
combination of Allele 1- and Allele 2-specific RFUs. This plot is an effective way to present
RFU data.
The calculation for normalized RFU follows the formulas presented in Livak et al. (1995).
Where:
• A
1
represents RFU for Allele 1
• A
2
represents RFU for Allele 2
• represents the mean RFU
NTC
A1 + A2
represents the sum of RFUs for the NTC sample of Allele 1 and Allele 2
Allelic Discrimination Spreadsheet
The Allelic Discrimination spreadsheet at the top right side of the Allelic Discrimination tab
shows the information shown in Table 32.
Custom Data View Tab
The Custom Data View tab simultaneously displays multiple panes in a customizable format
(Figure 59).
The Load a Preset View dropdown list offers a selection of display format templates. The
default view displayed is dependent on the file being analyzed. For example, if Melt Curve data
are present, the Amp+Melt default view is displayed.
The data view can be further customized by:
• Selecting an alternate preset view from the dropdown list
• Using the dropdown menu located at the top of an individual pane
• Using the Rows and Columns dropdown selection options
Table 32. Allelic Discrimination spreadsheet contents
Information Description
Well Well position in the plate.
RFU1 or C
q
1 RFU or C
q
for Allele1.
RFU2 or C
q
2 RFU or C
q
for Allele2.
Call Identity of the allele, including automatic Allele 1, Allele 2,
Heterozygote, None, Unknown, Control 1, Control 2.
Type Auto (Automatic) or Manual. Describes the way the call was
made. Automatic means the software selected the call.
Manual means the call was chosen by the user.
Normalized A
1
A
1
A
1
A
2
x NTC
A1 A2 +
 
+ +
-------------------------------------------------------------- =