通过二维天然-天然DNA琼脂糖凝胶电泳分析真核细胞的DNA结构

Analysis of DNA structures from eukaryotic cells by two-dimensional native-native DNA agarose gel electrophoresis.

作者信息

Ivessa Andreas S

机构信息

Department of Cell Biology and Molecular Medicine, University of Medicine and Dentistry of New Jersey, Newark, NJ, USA.

出版信息

Methods Mol Biol. 2013;1054:63-81. doi: 10.1007/978-1-62703-565-1_4.

DOI:10.1007/978-1-62703-565-1_4

PMID:23913285

Abstract

The neutral-neutral two-dimensional agarose gel technique is mainly used to determine the chromosomal positions where DNA replication starts, but it is also applied to visualize replication fork progression and breakage as well as intermediates in DNA recombination. Here we provide a step-by-step protocol to analyze the fairly underrepresented and fragile replication intermediates in yeast chromosomal DNA. The technique can also be adapted to analyze replication intermediates in chromosomal DNA of higher eukaryotic organisms.

摘要

中性-中性二维琼脂糖凝胶技术主要用于确定DNA复制起始的染色体位置，但也可用于观察复制叉的进展、断裂以及DNA重组中的中间体。在此，我们提供了一个逐步方案，用于分析酵母染色体DNA中相对较少且脆弱的复制中间体。该技术也可适用于分析高等真核生物染色体DNA中的复制中间体。