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Applied Biosystems 7500 Getting Started Guide

Applied Biosystems 7500
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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

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Applied Biosystems 7500 Specifications

General IconGeneral
Detection MethodFluorescence
Sample Capacity96-well plate
Thermal CyclerYes
Temperature Uniformity±0.5°C
TypeReal-Time PCR System
Thermal Accuracy±0.25°C
Detection Channels5
Weight45 kg
Power Requirements100-240 V, 50/60 Hz
Dynamic Range9 logs
Reaction Volume Range10-100 µL
Temperature Range4°C to 100°C
Thermal Cycler Block96-well
SoftwareSDS Software
DetectionMultiplex (up to 5 colors)

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