SpringerProtocols: Abstract: High-Throughput Real-Time PCR for Detection of Gene-Expression Levels

12. High-Throughput Real-Time PCR for Detection of Gene-Expression Levels

By: Bridget K. Wagner

, Zoltan Arany¹

Abstract

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While many high-throughput screening campaigns involve the measurement of protein levels or locations, at times it is desirable to measure the levels of gene expression in response to small molecules. Here, we describe a method for capturing mRNA in multiwell plates following compound treatment and measuring gene expression using real-time PCR. This streamlined protocol provides complementary information to conventional phenotypic cell-based assays, and is especially useful in cases where the gene of interest is thought to serve a regulatory function in downstream cellular phenotypes.

Affiliation(s): (1) Chemical Biology Program, Broad Institute of Harvard and MIT, Cambridge, MA 02142, USA
(2) Cardiovascular Institute at the Beth Israel Deaconess Medical Center Dana-Farber Cancer Institute, Boston, MA, USA

Book Title: Cell-Based Assays for High-Throughput Screening: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 486 | Year: 2009 | Page Range: 167-175 | DOI: 10.1007/978-1-60327-545-3_12

Subject: Biochemistry

Key Words: High-throughput gene expression - mRNA capture - Real-time PCR - Reverse transcription

References

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