通过变性梯度凝胶电泳检测突变

Detection of mutations by denaturing gradient gel electrophoresis.

作者信息

Børresen-Dale A L, Hovig E, Smith-Sørensen B

机构信息

The Norwegian Radium Hospital, Oslo, Norway.

出版信息

Curr Protoc Hum Genet. 2001 May;Chapter 7:Unit 7.5. doi: 10.1002/0471142905.hg0705s17.

DOI:10.1002/0471142905.hg0705s17

PMID:18428306

Abstract

This unit describes the procedure for determining the melting profile for a given PCR-amplified sequence by perpendicular denaturing gradient gel electrophoresis (DGGE) and, using that information, for developing a screening assay based on either parallel DGGE, CDGE (Constant Denaturant Gel Electrophoresis), or TTGE (Temporal/Temperature Gradient Electrophoresis). Four support protocols describe techniques for pouring perpendicular and parallel denaturing gradient gels, constant denaturant gels, and temporal temperature gradient gels.

摘要

本单元介绍了通过垂直变性梯度凝胶电泳（DGGE）确定给定PCR扩增序列的解链曲线的程序，并利用该信息开发基于平行DGGE、CDGE（恒定变性剂凝胶电泳）或TTGE（时间/温度梯度电泳）的筛选检测方法。四个支持方案描述了制备垂直和平行变性梯度凝胶、恒定变性剂凝胶以及时间温度梯度凝胶的技术。