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
