Format

Send to

Choose Destination
See comment in PubMed Commons below
Curr Protoc Chem Biol. 2012 Mar 1;4(1):49-63.

High-Throughput RT-PCR for small-molecule screening assays.

Author information

  • 1Chemical Biology Platform, Broad Institute of MIT and Harvard, Cambridge, MA 617-714-7373.

Abstract

Quantitative measurement of the levels of mRNA expression using real-time reverse transcription polymerase chain reaction (RT-PCR) has long been used for analyzing expression differences in tissue or cell lines of interest. This method has been used somewhat less frequently to measure the changes in gene expression due to perturbagens such as small molecules or siRNA. The availability of new instrumentation for liquid handling and real-time PCR analysis as well as the commercial availability of start-to-finish kits for RT-PCR has enabled the use of this method for high-throughput small-molecule screening on a scale comparable to traditional high-throughput screening (HTS) assays. This protocol focuses on the special considerations necessary for using quantitative RT-PCR as a primary small-molecule screening assay, including the different methods available for mRNA isolation and analysis.

KEYWORDS:

High Throughput Screening; Real-time PCR; gene expression; phenotypic screening; qRT-PCR

PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Wiley Icon for PubMed Central
    Loading ...
    Write to the Help Desk