Unit

UNIT 14.7 In situ Hybridization and Detection Using Nonisotopic Probes

  1. Joan H.M. Knoll1,
  2. Peter Lichter2

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1407s31

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Knoll, J. H. and Lichter, P. 2001. In situ Hybridization and Detection Using Nonisotopic Probes. Current Protocols in Molecular Biology. 14:14.7.

Author Information

  1. 1

    Harvard Medical School, Boston, Massachusetts

  2. 2

    Deutsches Krebsforschungszentrum, Heidelberg, Germany

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JUL 1995

This is not the most recent version of the article. View current version (1 JUL 2007)

Abstract

Nonisotopic in situ hybridization can be used to determine the cellular location and the relative levels of expression of specific transcripts within cells and tissues. RNA in prepared specimens is hybridized with a probe labeled nonisotopically with biotin or digoxigenin, which is generally detected by fluorescence or enzymatic methods. Fluorescence in situ hybridization (FISH), probably the most widely used method, is described in addition to techniques for amplification of weak fluorescent signals obtained in FISH. Nonisotopic probes can also be detected by enzymatic reactions using horseradish peroxidase or alkaline phosphatase, as described here.