The comet assay:: a method to measure DNA damage in individual cells
The comet assay:: a method to measure DNA damage in individual cells
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DOI：
10.1038/nprot.2006.5
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Banath, Judit P.
中科院分区：
文献类型：
--
作者：
Olive, Peggy L.;Banath, Judit P.
我们提出了彗星测定的程序，这是一种基于凝胶电泳的方法，可用于测量单个真核细胞中的DNA损伤。它用途广泛，操作相对简单，灵敏度高。尽管大多数研究利用其测量DNA单链断裂的能力，但对该方法的修改允许检测DNA双链断裂，交联，碱基损伤和细胞核凋亡。灵敏度的极限是每个二倍体哺乳动物细胞约50条链断裂。用酵母、原生动物、植物、无脊椎动物和哺乳动物制备的单细胞悬浮液中的DNA损伤及其修复也可以使用该试验进行研究。彗星测定法最初是为了测量哺乳动物细胞群体内DNA损伤和修复能力的变化而开发的，现在的应用范围从人类和哨兵动物生物监测（例如，在有毒废物场所爬行的蚯蚓的DNA损伤）到测量特定基因组序列的DNA损伤。该协议可在不到24小时内完成。
We present a procedure for the comet assay, a gel electrophoresis - based method that can be used to measure DNA damage in individual eukaryotic cells. It is versatile, relatively simple to perform and sensitive. Although most investigations make use of its ability to measure DNA single-strand breaks, modifications to the method allow detection of DNA double-strand breaks, cross-links, base damage and apoptotic nuclei. The limit of sensitivity is approximately 50 strand breaks per diploid mammalian cell. DNA damage and its repair in single-cell suspensions prepared from yeast, protozoa, plants, invertebrates and mammals can also be studied using this assay. Originally developed to measure variation in DNA damage and repair capacity within a population of mammalian cells, applications of the comet assay now range from human and sentinel animal biomonitoring ( e. g., DNA damage in earthworms crawling through toxic waste sites) to measurement of DNA damage in specific genomic sequences. This protocol can be completed in fewer than 24 h.