Chapter 5 Analyze the Relative Standard Curve Experiment
View the Raw Data Plot
109
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative C
T
Experiments
Notes
Analysis
Guidelines
When you analyze your own relative standard curve experiment, look for:
• Passive reference – The passive reference dye fluorescence level should be
relatively constant throughout the PCR process.
• Reporter dye – The reporter dye fluorescence level should display a flat region
corresponding to the baseline, followed by a rapid rise in fluorescence as the
amplification proceeds.
• Any irregularities in the signal – There should not be any spikes, dips, and/or
sudden changes in the fluorescence.
• Negative control wells – There should not be any amplification in the negative
control wells.
For More
Information
For more information on the Multicomponent Plot screen, open the 7500 Software Help
by clicking or pressing F1.
View the Raw Data Plot
The Raw Data Plot screen displays the raw fluorescence (not normalized) for each optical
filter for the selected wells during each cycle of the real-time PCR.
About the
Example
Experiment
In the relative standard curve example experiment, you review the Raw Data Plot screen
for a stable increase in signal (no abrupt changes or dips) from the appropriate filter.
View the Plot
1. In the navigation pane, select Analysis Raw Data Plot.
Note: If no data are displayed, click Analyze.
2. Display all 96 wells in the Raw Data Plot screen by clicking the upper left corner of
the plate layout in the View Plate Layout tab.
3. Click (Show a legend for the plot).
Note: This is a toggle button. When the legend is displayed, the button changes to
Hide the plot legend.
Note:
The legend displays the color code for each row of the reaction plate. In the
example shown below, Row A is red, Row B is yellow/green, Row C is green, and so on.
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