Applied Biosystems 7500 Getting Started Guide

Chapter 1 Introduction
About RQ Experiments
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 3
Notes
1
About RQ Experiments
RQ Experiment
Workflow
In this document, the term “RQ experiment” refers to the entire process of relative
quantification, beginning with generating cDNA from RNA (reverse transcription) and
ending with analyzing an RQ study. The RQ experiment workflow is shown on page iii.
RQ Studies with
the
7300/7500/7500
Fast System
The data-collection part of an RQ assay is a single-plate document, called the RQ Plate.
Amplification data from PCR runs is stored with sample setup information on the plate.
The data-analysis part of an RQ assay is a multi-plate document, called the RQ Study.
You can analyze up to ten RQ plates in a study. RQ Study documents neither control the
instrument, nor do they provide tools for setting up or modifying plates.
IMPORTANT! RQ Study software is an optional package for the 7300 instrument but it is
standard for the 7500 instrument and the 7500 Fast instrument.
The following figure illustrates the RQ Study process.
Note: The 7300/7500/7500 Fast system software uses the comparative method (∆∆C
T
)
to calculate relative quantities of a nucleic acid sequence. If you want to perform relative
quantification using the relative standard curve method, you should use an AQ assay
type and consult the Real-Time PCR Systems Chemistry Guide for details on how to set
up a run and analyze this type of assay.
RQ Study
Document
kidneyplate.sds
liverplate.sds
bladderplate.sds
7300/7500/
7500 Fast
S
y
stem
RQ Plate
Documents
Reaction
Plate
SDS
Software
SDS
Software
Bladder
Bladder
Bladder
Kidney
Kidney
Kidney
Liver
Liver
Liver

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)