abi 7500 - View the Amplification Plot

Chapter 9 Analyze the Comparative C
T
Experiment
View the Amplification Plot
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
166
Notes
View the Amplification Plot
The Amplification Plot screen displays amplification of all samples in the selected wells.
Three plots are available:
• ∆Rn vs Cycle – ∆Rn is the magnitude of normalized fluorescence generated by the
reporter at each cycle during the PCR amplification. This plot displays ∆Rn as a
function of cycle number. You can use this plot to identify and examine irregular
amplification and to view threshold and baseline values for the run.
• Rn vs Cycle – Rn is the fluorescence from the reporter dye normalized to the
fluorescence from the passive reference. This plot displays Rn as a function of cycle
number. You can use this plot to identify and examine irregular amplification.
• C
T
vs Well – C
T
is the PCR cycle number at which the fluorescence meets the
threshold in the amplification plot. This plot displays C
T
as a function of well
position. You can use this plot to locate outlying amplification (outliers).
Each plot can be displayed as a linear or log10 graph.
About the
Example
Experiment
In the comparative C
T
example experiment, you review each target in the Amplification
Plot for:
• Correct baseline and threshold values
•Outliers