Applied Biosystems 7500 Getting Started Guide

vi Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide
Chapter 5 Generating Data from RQ Plates – 7500 Fast System 37
Before You Begin . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Preparing the PCR Master Mix . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
Preparing the Reaction Plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40
Creating a Relative Quantification (RQ) Plate Document . . . . . . . . . . . . . . . . . . . . . 43
Specifying Thermal Cycling Conditions and Starting the Run . . . . . . . . . . . . . . . . . 48
Analyzing and Viewing RQ Plate Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Exporting RQ Plate Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
Chapter 6 Analyzing Data in an RQ Study 59
Creating an RQ Study Document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
Configuring Analysis Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63
Adjusting the Baseline and Threshold . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65
Analyzing and Viewing the Results of the RQ Study . . . . . . . . . . . . . . . . . . . . . . . . 70
Reanalyzing an RQ Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
Omitting Samples from a Study . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
Exporting RQ Study Data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
Appendix A Creating Detectors 79
Appendix B Example RQ Experiment 81
References 89
Index 91

Applied Biosystems 7500 Specifications

General

Detection Method	Fluorescence
Sample Capacity	96-well plate
Thermal Cycler	Yes
Temperature Uniformity	±0.5°C
Type	Real-Time PCR System
Thermal Accuracy	±0.25°C
Detection Channels	5
Weight	45 kg
Power Requirements	100-240 V, 50/60 Hz
Dynamic Range	9 logs
Reaction Volume Range	10-100 µL
Temperature Range	4°C to 100°C
Thermal Cycler Block	96-well
Software	SDS Software
Detection	Multiplex (up to 5 colors)