The Comet-FISH assay for the analysis of DNA damage and repair.
The Comet-FISH assay for the analysis of DNA damage and repair.
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用于分析 DNA 损伤和修复的 Comet-FISH 测定。
DOI：
10.1007/978-1-60761-789-1_9
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Spivak,Graciela
中科院分区：
文献类型：
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作者：
Spivak,Graciela
在本章中，我介绍了我们实验室使用的碱性单细胞凝胶电泳法(彗星试验)和荧光原位杂交(FISH)技术，以研究紫外线诱导的人类细胞损伤的发生和修复。彗星-FISH方法允许对整个基因组和特定染色体区域的DNA损伤及其修复进行同步和比较分析。这种非常敏感的方法可以应用于任何损伤，例如由化学致癌物和细胞代谢产物引起的损伤，这些产物可以转化为DNA单链或双链断裂。讨论了该方法在具体应用中的独特优势和局限性。
In this chapter, I describe the alkaline single-cell gel electrophoresis (Comet assay) combined with fluorescence in situ hybridization (FISH) technology, used in our laboratory, to study the incidence and repair of lesions induced in human cells by ultraviolet light. The Comet-FISH method permits the simultaneous and comparative analysis of DNA damage and its repair throughout the genome and in defined chromosomal regions. This very sensitive approach can be applied to any lesion, such as those induced by chemical carcinogens and products of cellular metabolism that can be converted to DNA single- or double-strand breaks. The unique advantages and limitations of the method for particular applications are discussed.