【摘要】 单细胞凝胶电泳技术又称为彗星实验,是最近几年发展起来的一种快速、简单、灵敏、可靠的检测细胞核DNA损伤的技术。总结了近几年来单细胞凝胶电泳技术的发展、原理、方法及其应用,并指出其下一步的发展趋势。彗星实验中,镶嵌于琼脂糖中的细胞核在电场中向正极移动,因细胞核与DNA片段迁移速率不同,而形成类似“彗星”的图像。目前采用的彗星实验有多种,可以检测诸如DNA双链断裂、单链断裂、碱不稳定位点等多种类型的DNA损伤。碱性彗星实验因其高灵敏度而被广泛采用。彗星实验的主要步骤包括细胞核悬浮液的获得、彗星电泳胶板制备、细胞裂解、DNA变性解旋、电泳、中和、染色和观察等。目前彗星实验广泛应用于各个研究领域,近年来开始用于环境污染的基因毒性研究和生物监测,并取得了迅速发展。更多还原

【Abstract】 The single cell gel electrophoresis technique (comet assay) is a simple,sensitive,reliable,and rapid method to detect DNA damage in eukaryotic cells.The paper reviewed the principles,advances and methodology of comet assay,and its applications in soil ecotoxicology,with its development in future predicted.Nuclei embedded in low-melting-point agrose in an agar gel sandwich migrated towards the anode under an electrical current with a different rate from that of DNA strands presented the appearance of a "comet".Different techniques are available with different sensitivity to detect DNA damages such as double strand breaks,single strand breaks,delay DNA repair sites and alkali labile sites.The basic procedure of the technique included cell suspension preparation,slide preparation,lysis,unwinding,electrophoresis,neutralization,dying and view.The method has been widely used in several areas in the past few years,especially in genotoxicology research in environmental biomonitoring.更多还原