Chapter 1 Get Started
About Standard Curve Experiments
7
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
Notes
About Standard Curve Experiments
Real-Time PCR
Experiments
Standard curve experiments are real-time PCR experiments. In real-time PCR
experiments:
• The instrument monitors the progress of the PCR as it occurs.
• Data are collected throughout the PCR process.
• Reactions are characterized by the point in time during cycling when amplification
of a target is first detected.
Note: In this guide, the term experiment refers to the entire process of performing a run
using the 7500/7500 Fast system, including setup, run, and analysis.
About Standard
Curve
Experiments
The standard curve method is used to determine the absolute target quantity in samples.
With the standard curve method, the 7500 software measures amplification of the target
in samples and in a standard dilution series. Data from the standard dilution series are
used to generate the standard curve. Using the standard curve, the software interpolates
the absolute quantity of target in the samples.
Components
The following components are required when setting up PCR reactions for standard
curve experiments:
• Sample – The sample in which the quantity of the target is unknown.
• Standard – A sample that contains known standard quantities; used in quantitation
experiments to generate standard curves.
• Standard dilution series – A set of standards containing a range of known
quantities. The standard dilution series is prepared by serially diluting standards.
• Replicates – The total number of identical reactions containing identical samples,
components, and volumes.
• Negative Controls – Wells that contain water or buffer instead of sample template.
No amplification of the target should occur in negative control wells.
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