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通过变性梯度凝胶电泳(DGGE)进行突变检测。

Mutation detection by denaturing gradient gel electrophoresis (DGGE).

作者信息

Fodde R, Losekoot M

机构信息

MGC-Department of Human Genetics, Sylvius Laboratorium, Leiden, The Netherlands.

出版信息

Hum Mutat. 1994;3(2):83-94. doi: 10.1002/humu.1380030202.

Abstract

The molecular analysis of genetic diseases relies on several technical approaches which allow genetic and physical mapping, characterization of the gene structure, expression studies, and identification of disease-causing mutations. Denaturing gradient gel electrophoresis (DGGE) allows the rapid screening for single base changes in enzymatically amplified DNA. The technique is based on the migration of double-stranded DNA molecules through polyacrylamide gels containing linearly increasing concentrations of a denaturing agent. In this review DGGE and the several modifications of the original protocol are presented. Moreover, its applications in human molecular genetics are summarized together with a preliminary comparison with other mutation detection technologies such as chemical cleavage, RNase protection, and single-strand conformation polymorphism.

摘要

遗传疾病的分子分析依赖于多种技术方法,这些方法可用于基因和物理图谱绘制、基因结构表征、表达研究以及致病突变的鉴定。变性梯度凝胶电泳(DGGE)可用于快速筛查酶促扩增DNA中的单碱基变化。该技术基于双链DNA分子在含有线性增加浓度变性剂的聚丙烯酰胺凝胶中的迁移。在本综述中,介绍了DGGE及其对原始方案的几种改进。此外,总结了其在人类分子遗传学中的应用,并与其他突变检测技术(如化学切割、核糖核酸酶保护和单链构象多态性)进行了初步比较。

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