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Applied Biosystems 7300 Getting Started Guide
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Chapter 5
Generating Data f
rom RQ Plates – 7500 Fast System
T
roubleshooting
58
Applied Biosystems 7300/7500
/7500 Fast Real-Time PCR S
ystem Relative Quantificati
on Getting Started Guide
Notes
F
AST
FA
S
T
F
AST
67
69
Table of Contents
Default Chapter
3
Relative Quantification Experiment Workflow
3
Table of Contents
5
How to Use this Guide
7
Preface
7
How to Obtain more Information
9
How to Obtain Support
10
Introduction
11
About Relative Quantification
12
About the 7300/7500/7500 Fast System
12
Chapter 1 Introduction
12
About RQ Experiments
13
Designing an RQ Experiment
17
Selecting the PCR Method
18
Chapter 2 Designing an RQ Experiment
18
Specifying the Components of an RQ Experiment
19
Selecting the Chemistry
21
Selecting One- or Two-Step RT-PCR
22
Choosing the Probes and Primers
24
Performing Reverse Transcription
27
Guidelines for Preparing RNA
28
Chapter 3 Performing Reverse Transcription
28
Converting Total RNA to Cdna
29
Chapter 4 Generating Data from RQ Plates - 7300 or Standard 7500 System
31
Before You Begin
32
Preparing the PCR Master MIX
32
Preparing the Reaction Plate
33
Creating a Relative Quantification (RQ) Plate Document
35
Specifying Thermal Cycling Conditions and Starting the Run
40
Analyzing and Viewing RQ Plate Data
43
Exporting RQ Plate Data
46
Chapter 5 Generating Data from RQ Plates - 7500 Fast System
47
Before You Begin
48
Preparing the PCR Master MIX
48
Preparing the Reaction Plate
50
Creating a Relative Quantification (RQ) Plate Document
53
Specifying Thermal Cycling Conditions and Starting the Run
58
Analyzing and Viewing RQ Plate Data
62
Exporting RQ Plate Data
65
Troubleshooting
66
Analyzing Data in an RQ Study
69
Creating an RQ Study Document
70
Chapter 6 Analyzing Data in an RQ Study
70
Configuring Analysis Settings
73
Adjusting the Baseline and Threshold
75
Analyzing and Viewing the Results of the RQ Study
80
Reanalyzing an RQ Study
84
Omitting Samples from a Study
85
Exporting RQ Study Data
88
Appendix A Creating Detectors
89
Appendix B Example RQ Experiment
92
References
100
Index
102
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Applied Biosystems 7300 Specifications
General
Brand
Applied Biosystems
Model
7300
Category
Laboratory Equipment
Language
English
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