检测细胞DNA断裂损伤效应的彗星实验法的改良Improvements of the Comet Assay for Rapid Detecting the Rupture and Damage of DNA in Cells
赖金龙;付倩;任莎;吴国;陶宗娅;张红;罗学刚;
摘要(Abstract):
为了解决彗星实验过程中常出现的脱胶、细胞核分离操作繁琐、重复性低等问题,对彗星实验方法进行了改良,初步建立了彗星实验的快速操作流程。结果显示,通过对载玻片进行预处理,可确保凝胶悬挂均匀;采用改良机械法分离的细胞核浓度适中;以0.5%(w/v)涂层琼脂糖作为基层、以1.5%(w/v)低熔点包埋琼脂糖作为叠加层的"双层凝胶法",辅以"推片法"铺胶,操作便捷且不发生脱胶现象;细胞核膜经裂解处理后再进行电泳和荧光观察,彗星图像清晰,杂质少。应用改良后的彗星实验方法,操作简便,耗时更短,实验效果良好,可快速检测出细胞DNA损伤效应。
关键词(KeyWords): 铯;蚕豆根尖细胞;遗传毒性;DNA损伤;彗星实验;改良
基金项目(Foundation): 国家核设施退役及放射性废物治理科研重点项目(14ZG6101);; 四川省教育厅重点课题(14ZA0030);; 四川省生物质改性材料工程技术研究中心(西南科技大学)开放课题(12zxbk03);; 四川师范大学校级青年课题(14qn07);; 四川师范大学实验技术课题(SYJS2014-14)
作者(Author): 赖金龙;付倩;任莎;吴国;陶宗娅;张红;罗学刚;
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