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Author:
Date:
2013-09-20
[Abstract] The Comet assay (or Single Cell Gel Electrophoresis assay) is a sensitive technique to detect DNA damage at the level of an individual cell. This technique is based on micro-electrophoresis of cells DNA content. Briefly, cells are embedded in agarose, lysed and submitted to an electric field, before the staining step with a fluorescent DNA binding dye. Damaged DNA (charged DNA) migrates in this field, forming the tail of a “comet”, while undamaged DNA remained in the head of the “comet”. The following document describes the protocol to realize a neutral comet assay. This assay can be applied to different cell types and has been useful for numerous applications in fields of toxicology or DNA damage and repair.
[摘要] 彗星测定（或单细胞凝胶电泳测定）是在单个细胞水平检测DNA损伤的敏感技术。 这种技术是基于细胞DNA含量的微电泳。 简言之，将细胞嵌入琼脂糖中，裂解并在用荧光DNA结合染料染色步骤之前将其送入电场。 损伤的DNA（带电DNA）在该领域迁移，形成“彗星”的尾部，而未损坏的DNA保留在“彗星”的头部。 以下文件描述了实现中性彗星测定的方案。 该测定可应用于不同的细胞类型，并且可用于毒理学或DNA损伤和修复领域的许多应用。
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