Schvartzman Jorge B, Martínez-Robles María-Luisa, Hernández Pablo, Krimer Dora B
Department of Cell and Molecular Biology, Centro de Investigaciones Biológicas (CSIC), Madrid, Spain.
Methods Mol Biol. 2013;1054:121-32. doi: 10.1007/978-1-62703-565-1_7.
Two-dimensional (2D) agarose gel electrophoresis is nowadays one of the best methods available to analyze DNA molecules with different masses and shapes. The possibility to use nicking enzymes and intercalating agents to change the twist of DNA during only one or in both runs, improves the capacity of 2D gels to discern molecules that apparently may look alike. Here we present protocols where 2D gels are used to understand the structure of DNA molecules and its dynamics in living cells. This knowledge is essential to comprehend how DNA topology affects and is affected by all the essential functions that DNA is involved in: replication, transcription, repair and recombination.
二维（2D）琼脂糖凝胶电泳是目前可用于分析具有不同质量和形状的DNA分子的最佳方法之一。在一次或两次电泳过程中使用切口酶和嵌入剂来改变DNA的扭曲度，提高了二维凝胶辨别看似相似分子的能力。在这里，我们展示了使用二维凝胶来理解DNA分子结构及其在活细胞中的动态变化的实验方案。这些知识对于理解DNA拓扑结构如何影响以及如何受到DNA所涉及的所有基本功能（复制、转录、修复和重组）的影响至关重要。
