Specifying Thermal Cycling Conditions And Starting The Run - Applied Biosystems 7500 Getting Started Manual

Chapter 4 Generating Data from RQ Plates – 7300 or Standard 7500 System
STANDARD STANDARD STANDARD

Specifying Thermal Cycling Conditions and Starting the Run

Specifying Thermal Cycling Conditions and Starting the Run
Default Thermal
Cycling
Conditions for
PCR
1) RT Step
10 min @ 25 ° C
AmpErase
2) PCR Step
2 min @ 50 ° C
Notes
30
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide
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)
HOLD
120 min @ 37 ° C
Initial Steps
®
UNG AmpliTaq Gold
Activation Polymerase Activation
HOLD
10 min @ 95 ° C
HOLD
®
DNA
HOLD
15 sec @ 95 ° C
* For reference only.
RT is complete at this point.
PCR (Each of 40 cycles)
Melt Anneal/
Extend
CYCLE
1 min @ 60 ° C