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
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
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