Unit

UNIT 7.8 Analysis of DNA Denaturation

  1. Zbigniew Darzynkiewicz,
  2. Gloria Juan

Published Online: 1 MAY 2001

DOI: 10.1002/0471142956.cy0708s03

Current Protocols in Cytometry

Current Protocols in Cytometry

How to Cite

Darzynkiewicz, Z. and Juan, G. 2001. Analysis of DNA Denaturation. Current Protocols in Cytometry. 3:7.8:7.8.1–7.8.9.

Author Information

  1. New York Medical College, Elmsford, New York

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: JAN 1998

Abstract

This unit provides a detailed method for evaluating in situ DNA denaturation by flow cytometry. The principal technique is based upon the metachromatic properties of acridine orange. This technique has a number of advantages over traditional biochemical methods, but requires very precise methodology; the result is excellent clarity and differentiation between single-stranded and double-stranded DNA. This unit provides full details on the correct use of acridine orange together with an excellent discussion of the pitfalls and problems.

Keywords:

  • flow cytometry;
  • DNA denaturation;
  • acridine orange;
  • metachromatic dyes;
  • differential staining;
  • single-stranded DNA;
  • double-stranded DNA