Applied Biosystems 7500 Fast Getting Started

Chapter 1 Get Started
About the 7500/7500 Fast System
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
2
Notes
About the 7500/7500 Fast System
The Applied Biosystems 7500/7500 Fast Real-Time PCR System is a 96-well, five-color
platform that uses fluorescence-based polymerase chain reaction (PCR) reagents to provide:
• Quantitative detection of target nucleic acid sequences (targets) using real-time
analysis.
• Qualitative detection of targets using post-PCR (endpoint) analysis.
• Qualitative analysis of the PCR product (achieved by melt curve analysis that occurs
post-PCR).
About Data
Collection
The 7500/7500 Fast system collects raw fluorescence data at different points during a
PCR, depending on the type of run that the instrument performs:
Regardless of the run type, a data collection point or read on the 7500/7500 Fast
instrument consists of three phases:
1. Excitation – The instrument illuminates all wells of the reaction plate within the
instrument, exciting the fluorophores in each reaction.
2. Emission – The instrument optics collect the residual fluorescence emitted from the
wells of the reaction plate. The resulting image collected by the device consists only
of light that corresponds to the range of emission wavelengths.
3. Collection – The instrument assembles a digital representation of the residual
fluorescence collected over a fixed time interval. The 7500 software stores the raw
fluorescence image for analysis.
After a run, the 7500 software uses region of interest (ROI), optical, dye, and background
calibrations to determine the location and intensity of the fluorescence in each read, the
dye associated with each fluorescent signal, and the significance of the signals.
Run Type Data Collection Point
Real-time runs Standard curve The instrument collects data after each extension
step of the PCR.
Relative standard curve
Comparative C
T
(∆∆C
T
)
Post-PCR
(endpoint) runs
Genotyping The instrument collects data:
• Before the PCR (For presence/absence
experiments, data collection before the PCR is
optional but recommended.)
• (Optional) During the PCR. The instrument can
collect data during the run (real-time);
collecting data during the run can be helpful
for troubleshooting.
•After the PCR.
Presence/absence

Applied Biosystems 7500 Fast Specifications

General

Brand	Applied Biosystems
Model	7500 Fast
Category	Laboratory Equipment
Language	English