Applied Biosystems 7500 Getting Started Guide

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

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)