abi 7500 User manual

Applied Biosystems
7500/7500 Fast
Real-Time PCR System
Relative Standard Curve and
Comparative CTExperiments
Getting Started Guide

Applied Biosystems
7500/7500 Fast
Real-Time PCR System
Relative Standard Curve and
Comparative CTExperiments
Get Started
Design the
Relative Standard
Curve Experiment
Prepare the
Relative Standard
Curve Reactions
Run the Relative
Standard Curve
Experiment
Analyze the
Relative Standard
Curve Experiment
Design the
Comparative CT
Experiment
Prepare the
Comparative CT
Reactions
Run the
Comparative CT
Experiment
Analyze the
Comparative CT
Experiment
Getting Started Guide

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Part Number 4387783 Rev. C
06/2010

Contents
iii
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Relative Standard Curve
and Comparative CTExperiments
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
How to Use This Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
How to Obtain More Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ix
How to Obtain Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xi
Safety Conventions Used in This Document . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xii
Symbols on Instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiii
Safety Labels on Instruments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xiv
General Instrument Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xvi
Chemical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xvii
Chemical Waste Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xix
Electrical Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xx
Physical Hazard Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xx
Biological Hazard Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxi
Workstation Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . xxi
Safety and Electromagnetic Compatibility (EMC) Standards . . . . . . . . . . . . . . . . . . . xxii
Chapter 1 Get Started. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
About the 7500/7500 Fast System . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Supported Consumables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
About Relative Standard Curve and Comparative CTExperiments . . . . . . . . . . . . . . . . 7
How to Use This Guide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
About the Example Experiments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Example Experiment Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Chapter 2 Design the Relative Standard Curve Experiment. . . . 19
Chapter Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Create a New Experiment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
Define the Experiment Properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Define the Methods and Materials . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
Set Up the Targets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Set Up the Standards . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Set Up the Samples . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Set Up the Relative Quantitation Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
Set Up the Run Method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
Review the Reaction Setup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36