Cell-Based Assays for High-Throughput Screening

Volume 486 of the series Methods in Molecular Biology pp 167-175


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

  • Bridget K. WagnerAffiliated withChemical Biology Program, Broad Institute of Harvard and MIT Email author 
  • , Zoltan AranyAffiliated withCardiovascular Institute at the Beth Israel Deaconess Medical Center Dana-Farber Cancer Institute

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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.

Key words

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