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天然/变性二维DNA电泳及其在重组中间体分析中的应用。

Native/Denaturing two-dimensional DNA electrophoresis and its application to the analysis of recombination intermediates.

作者信息

Lao Jessica P, Tang Shangming, Hunter Neil

机构信息

Howard Hughes Medical Institute and the Departments of Microbiology & Molecular Genetics, Molecular & Cellular Biology and Cell Biology & Human Anatomy, University of California Davis, Davis, CA, USA.

出版信息

Methods Mol Biol. 2013;1054:105-20. doi: 10.1007/978-1-62703-565-1_6.

Abstract

Two-dimensional (2D) gel electrophoresis employs distinct electrophoretic conditions to better resolve complex mixtures of molecules. In combination with Southern analysis, 2D agarose gel electrophoresis is routinely employed to detect and analyze DNA intermediates that arise during the replication and repair of chromosomes. By separating intermediates into their component single-strands, native/denaturing 2D gels can reveal structure that is not apparent under native conditions alone. Here, we describe a general method for native/denaturing two-dimensional gel electrophoresis and its application to understanding the DNA strand-composition of recombination intermediates formed during meiosis.

摘要

二维(2D)凝胶电泳采用不同的电泳条件,以更好地分离复杂的分子混合物。与Southern分析相结合,二维琼脂糖凝胶电泳通常用于检测和分析染色体复制和修复过程中出现的DNA中间体。通过将中间体分离成其组成单链,天然/变性二维凝胶可以揭示仅在天然条件下不明显的结构。在这里,我们描述了一种用于天然/变性二维凝胶电泳的通用方法及其在理解减数分裂过程中形成的重组中间体的DNA链组成方面的应用。

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