Chapter 6
Design the Comparative C
T
Experiment
Define the Methods and Materials
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
116
Notes
Define the Methods and Materials
In the Methods and Materials screen, select the quantitation method, reagents, ramp
speed, and PCR template to use for the experiment.
About the
Example
Experiment
In the comparative C
T
example experiment:
• The comparative C
T
quantitation method is used.
• TaqMan
®
reagents are used.
• The standard ramp speed is used in the instrument run.
• cDNA (prepared from total RNA isolated from liver, kidney, brain, and universal
tissues) is the template type. You must first perform reverse transcription to convert
the RNA to cDNA (see “Prepare the Template” on page 139).
Complete the
Methods and
Materials Screen
1.
Select
Comparative C
T
(
∆∆
C
T
)
as the quantitation method.
2.
Select
TaqMan
®
Reagents
for the reagents. (default)
IMPORTANT!
Fast runs cannot be performed on a 7500 system.
3.
Select
Standard (~ 2 hours to complete a run)
for the ramp speed.
4.
Select
cDNA (complementary DNA)
for the template type. (default)
5.
Click
Next
.
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