Unit

UNIT 7.3 Mismatch Detection Using Heteroduplex Analysis

  1. Anne-Lise Børresen

Published Online: 1 AUG 2002

DOI: 10.1002/0471142905.hg0703s33

Current Protocols in Human Genetics

Current Protocols in Human Genetics

How to Cite

Børresen, A.-L. 2002. Mismatch Detection Using Heteroduplex Analysis. Current Protocols in Human Genetics. 33:7.3:7.3.1–7.3.3.

Author Information

  1. The Norwegian Radium Hospital, Oslo, Norway

Publication History

  1. Published Online: 1 AUG 2002
  2. Published Print: APR 2002

Abstract

This protocol describes a technique that can be used to identify point mutations or single-base polymorphisms in heterozygous individuals. This technique takes advantage of the fact that heteroduplex molecules containing single-base mismatches can be separated under particular conditions of gel electrophoresis from nearly identical molecules containing no mismatches. RNA or DNA from a potentially heterozygous individual is amplified by PCR, and the products are denatured and allowed to renature, forming heteroduplexes. Renatured PCR products are run on nondenaturing mutation-detection-enhancement polyacrylamide gels (HydroLink MDE gels). On this gel, hybrid molecules containing a mismatch migrate more slowly than their corresponding homoduplexes. This protocol describes the analysis of unlabeled PCR products; however, radiolabeled PCR products can also be analyzed by this method.