Unit

UNIT 14.7 In Situ Hybridization and Detection Using Nonisotopic Probes

  1. Joan H.M. Knoll1,
  2. Peter Lichter2,
  3. Khldoun Bakdounes3,
  4. Isam-Eldin A. Eltoum3

Published Online: 1 JUL 2007

DOI: 10.1002/0471142727.mb1407s79

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Knoll, J. H., Lichter, P., Bakdounes, K. and Eltoum, I.-E. A. 2007. In Situ Hybridization and Detection Using Nonisotopic Probes. Current Protocols in Molecular Biology. 79:14.7:14.7.1–14.7.17.

Author Information

  1. 1

    University of Western Ontario, London, Canada

  2. 2

    Deutsches Krebsforschungszentrum, Heidelberg, Germany

  3. 3

    University of Alabama at Birmingham, Birmingham, Alabama

Publication History

  1. Published Online: 1 JUL 2007
  2. Published Print: JUL 2007

Abstract

Nonisotopic in situ hybridization can be used to determine the cellular location and relative levels of expression for specific transcripts within cells and tissues. RNA in specimen preparations is hybridized with a biotin- or digoxigenin-labeled probe, which is generally detected by fluorescence or enzymatic methods. Fluorescence in situ hybridization (FISH), probably the most widely used method, is described here, along with amplification of weak FISH signals. Nonisotopic probes can also be detected by enzymatic reactions using horseradish peroxidase or alkaline phosphatase, as described here. Curr. Protoc. Mol. Biol. 79:14.7.1-14.7.17. © 2007 by John Wiley & Sons, Inc.

Keywords:

  • nonisotopic detection;
  • in situ hybridization;
  • FISH;
  • enzymatic detection