Chapter 5 Generating Data from RQ Plates – 7500 Fast System
Creating a Relative Quantification (RQ) Plate Document
Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Relative Quantification Getting Started Guide 43
Notes
FAST
FAST
5
Creating a Relative Quantification (RQ) Plate Document
Overview An RQ Plate document stores data collected from an RQ run for a single plate. There
must be one RQ Plate document for every RQ plate. RQ Plate documents also store other
information, including sample names and detectors.
IMPORTANT! RQ plates run with standard thermal cycling conditions. Fast thermal
cycling conditions cannot be combined into a single RQ study.
Run Setup
Requirements
For each RQ plate document that you create, specify detectors, endogenous controls, and
detector tasks:
• A detector is a virtual representation of a gene-specific nucleic acid probe reagent
used in assays. You specify which detector to use for each target sequence.
Appendix A explains how to create detectors.
IMPORTANT! To conduct a comparative analysis of the data in a study, all the
plates in the study must contain a common set of detectors.
• An endogenous control(s) is defined in “Specifying the Components of an RQ
Experiment” on page 9. If your experiment consists of multiple plates, each plate
must have at least one endogenous control with at least three replicates. If your
experiment consists of a single plate with multiple samples, there must be an
endogenous control for each sample. All plates in an RQ experiment must use the
same endogenous control (for example, GAPDH).
• A detector task specifies how the software uses the data collected from the well
during analysis. You apply one of two tasks to each detector in each well of a plate
document.
Creating an RQ
Plate Document
You can enter sample information into a new plate document, import sample
information from existing plate documents, or use a template document to set up new
plate documents. This section describes setting up new plate documents. Refer to the
Online Help for information about importing sample information or using template
documents.
Task Symbol Apply to detectors of...
Target Wells that contain PCR reagents for the amplification of
target sequences.
Endogenous
Control
Wells that contain reagents for the amplification of the
endogenous control sequence.