abi 7500 Getting Started Manual
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Contents
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Chapter 6 Design the Comparative CT ExperimentSet Up the Targets119Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curveand Comparative CT ExperimentsNotesSet Up the TargetsIn the Targets screen, enter the number of targets you want to quantify in the PCRreaction plate, then set up the assay for each target.About theExampleExperimentIn the comparative CT example experiment:• Two targets are quantified in the reaction plate.• The Target 1 assay is set up for the endogenous control. For the example experimentthe target is human 18S. 18S serves as the endogenous control because its expressionlevels tend to be relatively stable.• The Target 2 through 6 assays are set up for the targets you are studying. For theexample experiment, the targets are EGR3, MAOB, OGDH, OSGEP, and SERPING1.Complete theTargets Screen1. Click the How many targets do you want to quantify in the reaction plate? field,then enter 6.Note: The number of rows in the target assays table is updated with the number youentered.2. Set up the Target 1 assay:a. Click the Enter Target Name cell, then enter 18S.b. In the Reporter drop-down list, select FAM (default).c. In the Quencher drop-down list, select NFQ-MGB (default).d. Leave the default in the Color field.3. Set up the remaining targets as shown below:4. Click Next.Note: For all targets, leave blank the (Optional) Enter Gene Name field. You can searchfor the gene/assay ID when you order your materials (see “Order Materials for theExperiment” on page 131).Target Target Name Reporter Quencher Color2 EGR3 FAM NFQ-MGB (Use default color)3 MAOB FAM NFQ-MGB (Use default color)4 OGDH FAM NFQ-MGB (Use default color)5 OSGEP FAM NFQ-MGB (Use default color)6 SERPING1 FAM NFQ-MGB (Use default color) |
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