Chapter 1 Get Started
About the Example Standard Curve Experiment
Applied Biosystems 7500/7500 Fast Real-Time PCR System Getting Started Guide for Standard Curve
Experiments
12
Notes
About the Example Standard Curve Experiment
To illustrate how to perform standard curve experiments, this guide leads you through the
process of designing, preparing, running, and analyzing an example experiment. The
example experiment represents a typical setup that you can use to quickly familiarize
yourself with a 7500/7500 Fast system.
Description
The objective of the standard curve example experiment is to determine the quantity of
the RNase P gene in two populations.
In the standard curve example experiment:
• The samples are genomic DNA isolated from two populations.
• The target is the RNase P gene.
• One standard curve is set up for the RNase P gene (target). The standard used for the
standard dilution series contains known quantities of the RNase P gene. Because a
single target is being studied, only one standard curve is required.
Note: In experiments where multiple targets are being studied, a standard curve is
required for each target.
• Three replicates of each sample and each dilution point in the standard curve are
performed to ensure statistical significance.
• The experiment is designed for singleplex PCR, where every well contains a
primer/probe set for a single target.
• Primer/probe sets are from Applied Biosystems RNase P assay.
Note: The human RNase P FAM
™
dye-labeled MGB probe is not available as a
TaqMan
®
Gene Expression Assay. It can be ordered as a Custom TaqMan
®
Gene
Expression Assay (PN 4331348).
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