Chapter 9 Analyze the Comparative C
T
Experiment
View the Multicomponent Plot
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
182
Notes
Analysis
Guidelines
When you analyze your own comparative C
T
experiment, carefully view the replicate
groups for outliers. If needed, remove outliers manually using the Well Table:
1. In the navigation pane, select Analysis Amplification Plot.
Note: If no data are displayed, click Analyze.
2. In the Plot Type drop-down list of the Amplification Plot screen, select C
T
vs Well.
3. Select the View Well Table tab.
4. In the Well Table:
a. In the Group By drop-down list, select Replicate.
b. Look for any outliers in the replicate group (be sure they are flagged).
c. Select the Omit check box next to the outlying well(s).
5. Click Analyze to reanalyze the experiment data with the outlying well(s) removed
from the analysis.
For More
Information
For more information on omitting wells from the analysis, open the 7500 Software Help
by clicking or pressing F1. Within the Help, search for the omit well topics:
1. Select the Search tab.
2. Enter omit well.
3. Click List Topics.
4. Double-click the topics you want to review.
View the Multicomponent Plot
The Multicomponent Plot screen displays the complete spectral contribution of each dye
in a selected well over the duration of the PCR.
About the
Example
Experiment
In the comparative C
T
example experiment, you review the Multicomponent Plot screen
for:
•ROX
™
dye (passive reference)
•FAM
™
dye (reporter)
• Spikes, dips, and/or sudden changes
• Amplification in the negative control wells
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