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General| Instrument Type | Real-Time PCR System |
|---|---|
| Detection Method | Fluorescence |
| Sample Capacity | 96-well plate |
| Temperature Accuracy | ±0.25°C |
| Temperature Uniformity | ±0.5°C |
| Number of Channels | 5 |
| Detection Channels | 5 |
| Manufacturer | Applied Biosystems |
| Thermal Range | 4°C to 100°C |
| Ramp Rate | Max 3.5°C/sec |
| Heating Rate | 3.5°C/sec |
| Cooling Rate | 2.5°C/sec |
| Software Version | v2.3 |
| Power Requirements | 100-240 V, 50/60 Hz |
Introduces the Applied Biosystems 7500/7500 Fast Real-Time PCR System and its capabilities.
Lists the consumables supported by the 7500 and 7500 Fast systems.
Explains relative standard curve and comparative CT (ΔΔCT) experiments in real-time PCR.
Explains how to use this guide as a tutorial and for performing own experiments.
Explains how to use the Design Wizard to set up the relative standard curve example experiment.
Describes how to create a new experiment using the Design Wizard in the 7500 software.
Enter identifying info, select instrument type, and experiment type.
Select quantitation method, reagents, ramp speed, and PCR template.
Enter number of targets and set up the assay for each target.
Enter number of points and replicates for standard curves.
Explains how to prepare PCR reactions for the relative standard curve example experiment.
Prepare template for PCR reactions using the High-Capacity cDNA Reverse Transcription Kit.
Perform sample dilutions before adding samples to the final reaction mix.
Prepare reaction mix using components and volumes calculated by the 7500 software.
Explains how to perform a run on the Applied Biosystems 7500/7500 Fast Real-Time PCR System.
Open example experiment file and load sealed reaction plate into the instrument.
Starts the 7500/7500 Fast instrument run after preparation.
View run progress in real time and check available plots for problems.
Explains how to review analyzed data and publish it, and troubleshoot questionable results.
Analyzes the experiment and displays results in analysis screens.
Displays the standard curve and calculates quantity of unknown target.
Displays amplification of all samples in selected wells, showing plots.
Explains how to use Design Wizard for comparative CT (ΔΔCT) example experiment setup.
Describes creating a new experiment using the Design Wizard in the 7500 software.
Enter identifying info, select instrument type, and experiment type for the study.
Select quantitation method, reagents, ramp speed, and PCR template.
Enter number of targets and set up the assay for each target.
Enter number of samples, replicates, and negative controls.
Explains how to prepare PCR reactions for the comparative CT example experiment.
Prepare template for PCR reactions using the High-Capacity cDNA Reverse Transcription Kit.
Perform sample dilutions before adding samples to the final reaction mix.
Prepare reaction mix using components and volumes calculated by the 7500 software.
Explains how to perform a run on the Applied Biosystems 7500/7500 Fast Real-Time PCR Systems.
Open example experiment file and load sealed reaction plate into the instrument.
Performs the run by following procedures from Chapter 4.
Explains how to review analyzed data, publish it, and troubleshoot results.
Analyzes the experiment and displays results in analysis screens.
Displays relative quantitation results in gene expression profile and well table.
Displays amplification of all samples in selected wells, showing plots.
Describes ways to publish experiment data, like saving or printing plots.
Displays software flags, frequency, and location for the open experiment.
Removes wells with insufficient amplification or outliers from analysis.
Displays spectral contribution of each dye in a selected well over PCR duration.
Displays raw fluorescence for each optical filter during each PCR cycle.
Combines analysis of experiments using the comparative CT method.
Sets up an experiment using Advanced Setup for custom design.
Runs reactions with no reaction plate setup information.
Uses a template to create new experiments with same setup information.
Sets up a new experiment using setup data exported from another experiment.
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