用于分离DNA拓扑异构体的二维凝胶电泳
Two-Dimensional Gel Electrophoresis to Resolve DNA Topoisomers.
作者信息
Gibson Elizabeth G, Oviatt Alexandria A, Osheroff Neil
机构信息
Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, TN, USA.
Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN, USA.
出版信息
Methods Mol Biol. 2020;2119:15-24. doi: 10.1007/978-1-0716-0323-9_2.
Abstract
Agarose gel electrophoresis is one of the most straightforward techniques that can be used to differentiate between topoisomers of closed circular DNA molecules. Generally, the products of reactions that monitor the interconversion of DNA between negatively supercoiled and relaxed DNA or positively supercoiled and relaxed DNA can be resolved by one-dimensional gel electrophoresis. However, in more complex reactions that contain both positively and negatively supercoiled DNA, one-dimensional resolution is insufficient. In these cases, a second dimension of gel electrophoresis is necessary. This chapter describes the technique of two-dimensional agarose gel electrophoresis and how it can be used to resolve a spectrum of DNA topoisomers.
摘要
琼脂糖凝胶电泳是可用于区分闭环DNA分子拓扑异构体的最直接技术之一。一般来说，监测DNA在负超螺旋与松弛DNA之间或正超螺旋与松弛DNA之间相互转化的反应产物，可通过一维凝胶电泳进行分离。然而，在同时含有正超螺旋和负超螺旋DNA的更复杂反应中，一维分辨率是不够的。在这些情况下，二维凝胶电泳是必要的。本章介绍二维琼脂糖凝胶电泳技术及其如何用于分离一系列DNA拓扑异构体。
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