View The Amplification Plot - Applied Biosystems 7500 Getting Started

Chapter 5 Analyze the Experiment

View the Amplification Plot

View the Amplification Plot

About the
Example
Experiment
View the
Amplification Plot
Notes
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Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
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 vs Well – C is the PCR cycle number at which the fluorescence meets the
T
T
threshold in the amplification plot. This plot displays C
position. You can use this plot to locate outlying amplification (outliers).
Each plot can be displayed on a linear or log10 scale.
In the standard curve example experiment, you review the target in the Amplification
Plot screen for:
• Correct baseline and threshold values
• Outliers
1. In the navigation pane, select Analysis
If no data are displayed, click Analyze.
Note:
2. Display the RNase P wells in the Amplification Plot screen:
Select the View Plate Layout tab.
a.
In the Select Wells With drop-down lists, select Target, then RNase P
b.
TAMRA.
2a
3. In the Amplification Plot screen:
In the Plot Type drop-down list, select ∆Rn vs Cycle (default).
a.
In the Plot Color drop-down list, select Well (default).
b.
as a function of well
T
Amplification Plot.
2b
Experiments