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
30 Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide
Notes
STANDARD
STANDARD
STANDARD
Specifying Thermal Cycling Conditions and Starting the Run
Default Thermal
Cycling
Conditions for
PCR
If you selected the two-step RT-PCR method for your RQ experiment (recommended),
you have already completed the RT step and are ready to PCR amplify cDNA.
The default thermal cycling conditions for the PCR step of the procedure, shown in the
following table, should appear in the Instrument tab.
Times and Temperatures (Two-step RT-PCR)
1) RT Step
HOLD HOLD * For reference only.
RT is complete at this point.
10 min @ 25 ° C 120 min @ 37 ° C
2) PCR Step
Initial Steps PCR (Each of 40 cycles)
AmpErase
®
UNG
Activation
AmpliTaq Gold
®
DNA
Polymerase Activation
Melt Anneal/
Extend
HOLD HOLD CYCLE
2 min @ 50 ° C 10 min @ 95 ° C 15 sec @ 95 ° C1 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)