Applied Biosystems 7500 Getting Started Guide

Chapter 4 Generating Data from RQ Plates – 7300 or Standard 7500 System
Specifying Thermal Cycling Conditions and Starting the Run
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 31
Notes
STANDARD
STANDARD
STANDARD
4
Thermal Cycling
Conditions for
One-Step RT-PCR
If you select the one-step RT-PCR method, cDNA generation and amplification take
place simultaneously at this point in the workflow.
The following table shows the thermal cycling conditions for one-step RT-PCR
experiments.
Note: Refer to the Online Help for instructions on modifying thermal cycling
parameters.
Times and Temperatures (One-step RT-PCR)
Initial Steps PCR (Each of 40 Cycles)
Reverse Transcription AmpliTaq
®
Gold DNA
Polymerase Activation
Melt Anneal/Extend
HOLD HOLD CYCLE
30 min @ 48 ° C 10 min @ 95 ° C 15 sec @ 95 ° C 1 min @ 60 ° C

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)