Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 23)

Section 2 Quick Guides
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• 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
3. Click Relative Quantity (dCt).
4. Click Recalculate to see your results.
5. Relative Quantity results are graphed. The Data Table spreadsheet is accessed by
clicking Data Table. The Data Table spreadsheet lists the Condition and Gene name,
calculated expression values and C
T
values. Right-click to print or export this data to
Excel.
NOTE: Click the Settings tab, then select Gene List to enter a specific user-defined gene
reaction efficiency.
NOTE: Click the Settings tab, then select Condition List to select a particular sample as a
control sample.
If you need to compare these data to results obtained in other .opd files, you will need to enable
this file for Multi-file Gene Expression analysis, also called a Gene Study. To enable your file for
Gene Study:
1. Click the Enable for Gene Study button.
2. Go to the File menu to save your file.
Calculating Normalized Expression (ddC
T
, ∆∆C
T
)
To calculate Normalized Expression (ddC
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