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Normalizing Genetic Reporter Assays: Approaches and
Considerations for Increasing Consistency and Statistical Significance
Trista Schagat, Ph.D., Aileen Paguio, M.S., and
Kevin Kopish, B.S., Promega Corporation
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.
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