Gene Expression Analysis
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Plate Setup for Gene Expression Analysis
To perform gene expression analysis, the contents of the wells must include the following:
•Two or more targets. The two targets that represent different amplified genes or
sequences in your samples
• One or more reference targets. At least one target must be a reference target for
normalized expression. Assign all reference targets in the Experiment Settings window
(page 48) to analyze the data in Normalized Expression mode (ΔΔC(t)). Experiments that
do not contain a reference must be analyzed using Relative Expression mode (ΔC(t))
• Common samples. Your reactions must include common samples (minimum of two
required) to view your data plotted in the Gene Expression tab. These samples represent
different treatments or conditions for each of your target sequences. Assign a control
sample (optional) in the Experiment Settings window (page 48)
The requirements for Gene Expression setup in the Plate Editor depend on whether reaction
contents are singleplex PCR with one fluorophore in the reactions, or multiplex PCR with
more than one fluorophore in the reactions.
Figure 98 shows an example of the minimum contents of the wells for a singleplex gene
expression experiment.
Figure 98. Example of well contents in a singleplex gene expression experiment.
Figure 99 shows an example of the minimum contents of the wells for a multiplex gene
expression experiment.
Figure 99. Example of well contents in a multiplex gene expression experiment.
Gene Expression Tab
The Gene Expression tab in the Data Analysis window shows the relative expression of targets
in these two views:
• Gene Expression chart. Shows the real-time PCR data as normalized expression
(ΔΔC(t)) or relative quantity (ΔC(t))
• Spreadsheet. Shows a spreadsheet of the gene expression data
TIP: Right-click any chart or spreadsheet for options. Click the View/Edit Plate
button to open the Plate Editor, and change well contents in the plate.
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