Chapter 6 Design the Comparative C
T
Experiment
Review the Reaction Setup
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
128
Notes
c. Review the components and calculated volumes for the PCR reactions:
7. Repeat steps 2 through 6 for the remaining targets listed in the Selected Targets pane
(EGR3, MAOB, OGDH, OSGEP, and SERPING1).
Complete the Sample Dilution Calculations Tab
1. Select the Sample Dilution Calculations tab.
2. Click the Diluted Sample Concentration (10✕ for Reaction Mix) field, then
enter 50.0.
3. In the unit drop-down list, select ng/µL (default).
Component Volume (µL) for 1 Reaction
Master Mix (2.0✕) 25.00
Assay Mix (20.0✕)2.50
Sample (10✕)5.00
‡
‡ The sample volume is limited to 10% of the total reaction volume.
H2O 17.50
Total Volume 50.00
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6b
6a
6c
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