Unit

UNIT 4.5 High-Resolution FISH Analysis

  1. Henry H.Q. Heng1,
  2. Bradford Windle2,
  3. Lap-Chee Tsui3

Published Online: 1 FEB 2005

DOI: 10.1002/0471142905.hg0405s44

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Heng, H. H., Windle, B. and Tsui, L.-C. 2005. High-Resolution FISH Analysis. Current Protocols in Human Genetics. 44:4.5:4.5.1–4.5.23.

Author Information

  1. 1

    Center for Molecular Medicine and Genetics Wayne State University School of Medicine, Detroit, Michigan

  2. 2

    Virginia Commonwealth University, Richmond, Virginia

  3. 3

    University of Hong Kong, Hong Kong

Publication History

  1. Published Online: 1 FEB 2005
  2. Published Print: JAN 2005

Abstract

Map order, orientation, and gap or overlap distance of closely linked DNA probes may be determined using fluorescent hybridization to decondensed DNA. The linear arrangement of released chromatin fibers not only simplifies the task of gene ordering, but also provides higher resolution with probes separated by greater distances than can be achieved in FISH with intact interphase nuclei. The Basic Protocol 1 of this unit describes an alkaline lysis procedure for generating free chromatin from cultured cells for FISH analysis. A support protocol describes an empirical approach to optimize conditions for preparation of free chromatin. An Alternate Protocol 1 provides a method for producing free chromatin from cultured lymphocytes with drug treatment. The Basic Protocol 2, high-resolution FISH mapping with free chromatin, is a modification of the method used for FISH mapping of interphase nuclei.