UNIT 7.3 Mismatch Detection Using Heteroduplex Analysis
Published Online: 1 AUG 2002
Copyright © 2001 John Wiley & Sons, Inc. All rights reserved.
Lab Protocol Title
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.
- Published Online: 1 AUG 2002
- Published Print: APR 2002
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.