About The 7500/7500 Fast System - Applied Biosystems 7500 Getting Started
Real-time pcr system
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- Getting started manual (106 pages) ,
- Installation and maintenance manual (174 pages) ,
- Getting started manual (78 pages)
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Chapter 1 Get Started
About the 7500/7500 Fast System
About the 7500/7500 Fast System
About Data
Collection
Notes
2
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
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).
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:
Run Type
Real-time runs
Standard curve
Relative standard curve
Comparative C
Post-PCR
Genotyping
(endpoint) runs
Presence/absence
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.
Data Collection Point
The instrument collects data after each extension
step of the PCR.
(∆∆C
)
T
T
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.
Experiments
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