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Normalizing Genetic Reporter Assays Approaches and Considerations for Increasing Consistency and Statistical Significance

Trista Schagat, Aileen Paguio and Kevin Kopish
Promega Corporation
Publication Date: 2007

Abstract

Genetic reporter assays are used to study DNA sequences and cellular processes that control gene expression. In a typical reporter assay, cells are transfected with a vector that contains the sequence of interest cloned upstream of a reporter protein-coding sequence. Reporter activity is used as an indicator of the ability of the test sequence to regulate gene expression under the experimental conditions. Reporter activity is compared between different vector constructs (e.g., deletion analysis of promoter sequences; see reference 1) or treatment conditions (e.g., screening for G-protein-coupled receptor pathway modulators; see reference 2). Because variables such as cell number and transfection efficiency can have an unwanted effect on the magnitude of reporter expression, reporter data should be normalized. This article discusses approaches for normalization and highlights some key considerations for successful data analysis.

Cell Notes 17, 9–12.