Mettrick Karla A, Weaver Georgia M, Grainge Ian
School of Environmental and Life Sciences, University of Newcastle, Callaghan, NSW, Australia.
Methods Mol Biol. 2020;2119:61-72. doi: 10.1007/978-1-0716-0323-9_5.
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.
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。
