UNIT 14.7 In situ Hybridization and Detection Using Nonisotopic Probes
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- Published Print: JUL 1995
This is not the most recent version of the article. View current version (1 JUL 2007)
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.