Kohen R, Szyf M, Chevion M
Anal Biochem. 1986 May 1;154(2):485-91. doi: 10.1016/0003-2697(86)90019-9.
This communication describes a rapid and convenient procedure for quantitation of strand breaks in bacterial DNA, both in vitro and in vivo, using agarose gel electrophoresis. The electrophoretic determination of single strand breaks is carried out in alkaline medium, followed by renaturation of the gel and intercalation of the fluorescent dye, ethidium bromide. Double-strand breaks are determined by electrophoresis in neutral medium containing the dye. The distribution of DNA fragment sizes, the determination of the number-average molecular weight, the quantitation of the average number of DNA breaks per molecule, and the ratio between the single- and double-strand breaks are evaluated from microdensitometric scanning of the gels. The application of this analysis to damage caused by a combination of ascorbate and copper is demonstrated.
本通讯描述了一种快速便捷的程序，用于在体外和体内使用琼脂糖凝胶电泳对细菌DNA中的链断裂进行定量分析。单链断裂的电泳测定在碱性介质中进行，随后凝胶复性并插入荧光染料溴化乙锭。双链断裂通过在含有该染料的中性介质中进行电泳来测定。从凝胶的显微密度扫描评估DNA片段大小的分布、数均分子量的测定、每个分子中DNA断裂平均数量的定量以及单链和双链断裂之间的比例。证明了该分析方法在抗坏血酸盐和铜联合造成的损伤中的应用。
