abi 7500 Getting Started Manual
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Contents
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Chapter 5 Analyze the Relative Standard Curve ExperimentView the Raw Data Plot109Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curveand Comparative CT ExperimentsNotesAnalysisGuidelinesWhen you analyze your own relative standard curve experiment, look for:• Passive reference – The passive reference dye fluorescence level should berelatively constant throughout the PCR process.• Reporter dye – The reporter dye fluorescence level should display a flat regioncorresponding to the baseline, followed by a rapid rise in fluorescence as theamplification proceeds.• Any irregularities in the signal – There should not be any spikes, dips, and/orsudden changes in the fluorescence.• Negative control wells – There should not be any amplification in the negativecontrol wells.For MoreInformationFor more information on the Multicomponent Plot screen, open the 7500 Software Helpby clicking or pressing F1.View the Raw Data PlotThe Raw Data Plot screen displays the raw fluorescence (not normalized) for each opticalfilter for the selected wells during each cycle of the real-time PCR.About theExampleExperimentIn the relative standard curve example experiment, you review the Raw Data Plot screenfor 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 ofthe 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 toHide the plot legend.Note: The legend displays the color code for each row of the reaction plate. In theexample shown below, Row A is red, Row B is yellow/green, Row C is green, and so on. |
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