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细菌雷达检测法:细胞内共价复合物的分离与检测

Bacterial RADAR Assay: Isolation and Detection of Intracellular Covalent Complexes.

作者信息

Aldred Katie J

机构信息

Department of Biology, University of Evansville, Evansville, IN, USA.

出版信息

Methods Mol Biol. 2025;2928:17-23. doi: 10.1007/978-1-0716-4550-5_2.

Abstract

The bacterial rapid approach to DNA adduct recovery (RADAR) assay allows for the rapid lysis and trapping of DNA-topoisomerase cleavage complexes, as well as other proteins covalently attached to DNA. These covalently attached proteins can be quantified when the isolated DNA is immunoblotted. Significantly, the bacterial RADAR assay described here requires no large, expensive lab equipment, thus making it accessible to most laboratories. Furthermore, it does not require any long centrifugation steps, which allows for a relatively large number of samples to be processed in 1 day.

摘要

细菌快速DNA加合物回收(RADAR)分析方法能够快速裂解并捕获DNA-拓扑异构酶切割复合物,以及其他与DNA共价连接的蛋白质。当对分离出的DNA进行免疫印迹时,这些共价连接的蛋白质可以被定量。重要的是,本文所述的细菌RADAR分析方法不需要大型、昂贵的实验室设备,因此大多数实验室都可以采用。此外,该方法不需要任何长时间的离心步骤,这使得在一天内能够处理相对大量的样本。

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