Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 91)

Section 6 Data Analysis Module
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• Number of cells from which nucleic acid was isolated. Rel Quant/number of cells
represented in each sample
• Mass of tissue from which nucleic acid was isolated. Rel Quant/mass of tissue
represented in each sample
To calculate Relative Quantity (dC
T
):
1. In the PCR Quant screen with a .opd data file open, assess threshold and baseline
information for the data file and make changes if necessary.
2. Click the Gene Expr tab.
3. Make any changes to Gene and Condition (for example, Sample and Treatment)
assignments in the gene expression plate interface.
• Expand the gene expression plate interface view by clicking on the “+” button to
make well identification and selection easier. Highlight the wells in the gene
expression plate interface you wish to edit.
• Change the gene assignments in these wells by typing your desired name into the
gene pull-down men, then click enter to apply the name to the selected wells.
• Change the condition assignments in these wells by typing your desired name into
the condition pull-down menu, then click enter to apply the name to the selected
wells.
NOTE: The Gene and Condition Names have a character entry limit of 15 characters
• Minimize the gene expression plate interface by clicking on the “-” button to return to
the standard view of the Gene Expr tab window.
4. Click Relative Quantity (dCt).
5. Click Recalculate to see your results.
6. Relative quantity results are graphed. The Data Table spreadsheet is accessed by clicking
Data Table. The Data Table spreadsheet lists the Condition and Gene names, and
calculated expression values and C
T
values. Right click to print or export this data to
Excel.
NOTE: Use the Settings tab, then click Gene List to enter a specific user-defined gene
reaction efficiency.
NOTE: Use the Settings tab, then click Condition List to select a particular sample as
a control sample.
6.9.3 Specifying Gene and Condition Labels for Gene Expression
Analysis
Use the gene expression plate interface to edit gene names (for example actin, GAPDH), and
experimental conditions names (for example, treatment types, time course points, etc.). Use the
Settings option to select which of these genes and/or conditions to use for normalization and set
other analysis and display parameters.
Using the Gene Expression Plate Interface to Edit Gene and Condition Names
Open a .opd Real-Time PCR Data file.