用于可视化大肠杆菌DNA复制结构的中性-中性二维琼脂糖凝胶电泳

Neutral-Neutral 2-Dimensional Agarose Gel Electrophoresis for Visualization of E. coli DNA Replication Structures.

作者信息

Mettrick Karla A, Weaver Georgia M, Grainge Ian

机构信息

School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, Australia.

出版信息

Methods Mol Biol. 2020;2119:61-72. doi: 10.1007/978-1-0716-0323-9_5.

DOI:10.1007/978-1-0716-0323-9_5

PMID:31989514

Abstract

Neutral-neutral 2-dimensional agarose gel electrophoresis enables the detection of replication intermediate structures in DNA. Here we describe how DNA from Escherichia coli cells can be purified to retain replication intermediates and then be separated by size and shape using two consecutive agarose gel electrophoresis protocols. The DNA structures present within a localized region can be visualized by a Southern blotting/radioactive hybridisation protocol.

摘要

中性-中性二维琼脂糖凝胶电泳能够检测DNA中的复制中间结构。在此，我们描述了如何纯化来自大肠杆菌细胞的DNA以保留复制中间结构，然后使用两个连续的琼脂糖凝胶电泳方案按大小和形状进行分离。局部区域内存在的DNA结构可通过Southern印迹/放射性杂交方案进行可视化。