Set Up The Samples - abi 7500 Getting Started Manual

Set Up the Samples

In the Samples screen, enter the number of samples, replicates, and negative controls to
include in the reaction plate, enter the sample names, then select the sample/target
reactions to set up.
About the
In the comparative C
Example
• Four samples are used: cDNA prepared from total RNA isolated from four tissues
Experiment
• Four technical replicates are used. The replicates are identical reactions, containing
• No negative controls are used. Negative control reactions contain water instead of
Complete the 1. Click the How many samples do you want to test in the reaction plate? field, then
Samples Screen
2. Click the How many replicates do you need? field, then enter 4.
3. Click the How many negative controls do you need for each target assay? field,
4. Set up Sample 1:
5. Set up the remaining samples as shown below:
6. Select No Biological Replicates.
7. Select All Sample/Target Reactions to test all targets in all samples.
Notes
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C Experiments
T
example experiment:
T
(brain, kidney, liver, and universal). The samples contain unknown quantities of
EGR3, MAOB, OGDH, OSGEP, and SERPING1 (targets), and 18S (endogenous
control).
identical reaction components and volumes.
sample and should not amplify. The software automatically includes three negative
controls for each target assay.
Although the example experiment does not use negative controls,
IMPORTANT!
Applied Biosystems recommends them.
enter 4.
Note: The number of rows in the samples table is updated with the number you entered.
then enter 0.
Click the Enter Sample Name field, then enter Brain.
a.
Leave the default in the Color field.
b.
Sample Name
Kidney (Use default color)
Liver (Use default color)
Universal (Use default color)
Chapter 6 Design the Comparative C
Color
Experiment
T

Set Up the Samples

121