Speit Günter, Rothfuss Andreas
Institut für Humangenetik, Universität Ulm, Ulm, Germany.
Methods Mol Biol. 2012;920:79-90. doi: 10.1007/978-1-61779-998-3_6.
The comet assay (single-cell gel electrophoresis) is a simple and sensitive method for studying DNA damage and repair. In this microgel electrophoresis technique, a small number of cells suspended in a thin agarose gel on a microscope slide is lysed, electrophoresed, and stained with a fluorescent DNA-binding dye. Cells with increased DNA damage display increased migration of chromosomal DNA from the nucleus towards the anode, which resembles the shape of a comet. The assay has manifold applications in fundamental research for DNA damage and repair, in genotoxicity testing of novel chemicals and pharmaceuticals, environmental biomonitoring, and human population monitoring. This chapter describes a standard protocol of the alkaline comet assay and points to some useful modifications.
彗星试验（单细胞凝胶电泳）是一种用于研究DNA损伤与修复的简单且灵敏的方法。在这种微凝胶电泳技术中，将悬浮于显微镜载玻片上薄琼脂糖凝胶中的少量细胞进行裂解、电泳，并用一种荧光DNA结合染料染色。DNA损伤增加的细胞会显示出染色体DNA从细胞核向阳极迁移增加，这类似于彗星的形状。该试验在DNA损伤与修复的基础研究、新型化学品和药物的遗传毒性测试、环境生物监测以及人群监测等方面有多种应用。本章描述了碱性彗星试验的标准方案，并指出了一些有用的改进方法。
