Relative Quantification Experiment Workflow - Applied Biosystems 7500 Getting Started Manual

Chapter 1
Introduction
Designing
Chapter 2
an RQ
Experiment
Performing
Chapter 3 Primer Extended on mRNA
5´ 3´ Reverse
5´ cDNA
Primer Reverse
Oligo d(T) or random hexamer
Synthesis of 1st cDNA strand
3´ 5´ cDNA
Transcription
Generating
Data from
Chapter 4
STANDARD STANDARD
RQ Plates
Standard
(OR)
Generating
Data from
Chapter 5
FAST FAST
RQ Plates
Fast
Performing
Chapter 6
an RQ Study
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide
About the
About Relative
7300/7500/7500 Fast
Quantification
system
Specify the
Select the
components of
PCR method
an RQ experiment
Adjust RNA
Isolate total RNA
concentration
Prepare the Prepare the
PCR Master Mix reaction plate RQ Plate document
Create a new Configure
RQ Study document analysis settings
About
RQ Experiments
Select the Select one-step or
chemistry two-step RT-PCR
Convert
total RNA to cDNA
Program the
Create a new
thermal cycling
conditions
Adjust the
Analyze and
baseline and
view results
threshold

Relative Quantification Experiment Workflow

Choose probes
and primers
Save the
RQ Plate Start the run
RQ plate data
document
Export AQ
If necessary,
Plate document,
omit samples
if desired
Primer Extended on mRNA
5´ 3´
5´ cDNA
Primer Reverse
Oligo d(T) or random hexamer
Synthesis of 1st cDNA strand
3´ 5´ cDNA
STANDARD STANDARD
View
FAST FAST
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