Abstract
The External RNA Control Consortium (ERCC) is an ad-hoc group with approximately 70 members from private, public, and academic organizations. The group is developing a set of external RNA control transcripts that can be used to assess technical performance in gene expression assays. The ERCC is now initiating the Testing Phase of the project, during which candidate external RNA controls will be evaluated in both microarray and QRT-PCR gene expression platforms. This document describes the proposed experiments and informatics process that will be followed to test and qualify individual controls. The ERCC is distributing this description of the proposed testing process in an effort to gain consensus and to encourage feedback from the scientific community. On October 4–5, 2005, the ERCC met to further review the document, clarify ambiguities, and plan next steps. A summary of this meeting and changes to the test plan are provided as an appendix to this manuscript.
Highlights
External RNA controls consortium The External RNA Control Consortium (ERCC) is an adhoc group with approximately 70 members from private, public, and academic organizations
Protocols for the use of external RNA controls in clinical applications are included in the Molecular Methods 16-P document from the Clinical and Laboratory Standards Institute, and were developed in a formal, accredited, open, consensus forum including several ERCC members
The analysis approach was developed in a public workshop at National Institute of Standards and Technology (NIST) in June 2004, and is based upon the measurement of pooled transcripts at known concentrations
Summary
The microarray testing phases are designed to evaluate the candidate external RNA controls on both one-color and two-color platforms using the same pools of transcripts. During the Functional Testing phase, the target metrics determined during the Proof of Concept phase will be used to create a single-array DRC external RNA control pool. Repeat experiments in different labs Tasks for QRT-PCR subgroup The QRT-PCR testing phases are designed to use the same pools of transcripts developed for microarray testing and to evaluate the candidate external RNA controls on multiple QRT-PCR platforms. Compare microarray and QRT-PCR platforms Assay performance of a set of selected external RNA transcripts or pools with the same or different concentrations will be evaluated by microarray and QRT-PCR. Test plan tasks for ERCC Identify least burdensome path for collection and distribution of data
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