abi 7500 - Section Overview

Chapter 9 Analyze the Comparative C
T
Experiment
Section Overview
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
188
Notes
Section Overview
The 7500 software can combine the analysis of experiments that use the comparative C
T
method (∆∆C
T
) of relative quantitation.
For more information about methods of calculating relative quantitation, refer to the
Real-Time PCR Systems Reagent Guide.
About the
Example Study
In this section, you use two files:
• In the first half, you create a comparative C
T
example study file that contains setup
data, then save it to the experiments folder on your computer.
• In the second half, you view results in a example study file that is installed with the
7500 software. You can find the data file for the example study on your computer at:
<drive>:\Applied Biosystems\<software name>\experiments\
Comparative Ct Study Example.edm
where:
– <drive> is the computer hard drive on which the 7500 software is installed.
– <software name> is the current version of the 7500 software.
Note: The Comparative Ct Study Example.edm file is not designed to demonstrate the
use of biological replicate groups; an additional example study provided with the 7500
software provides a better example. You can find the alternate study file on your
computer at: <drive>:\Applied Biosystems\<software name>\experiments\
Comparative Ct Study Example (Biological Groups).edm.
In a comparative C
T
study, you can... You cannot...
• Specify the endogenous control(s) and
reference sample for the study.
• Select the control type when applicable.
• Set baseline and threshold values and
confidence levels, or set the number of
standard deviations for Comparative C
T
Min/Max.
• Omit wells individually or together through
their association with replicate groups
(technical or biological).
• Create, add, or modify samples.
• Create, add, or modify targets.
• Change assay tasks.
Note: You can perform these operations in
comparative C
T
experiments.