用于分析DNA损伤与修复的彗星荧光原位杂交检测法。

The Comet-FISH assay for the analysis of DNA damage and repair.

作者信息

Spivak Graciela

机构信息

Department of Biology, Stanford University, Stanford, CA, USA.

出版信息

Methods Mol Biol. 2010;659:129-45. doi: 10.1007/978-1-60761-789-1_9.

DOI:10.1007/978-1-60761-789-1_9

PMID:20809308

Abstract

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.

摘要

在本章中，我将描述我们实验室所使用的碱性单细胞凝胶电泳（彗星试验）与荧光原位杂交（FISH）技术相结合的方法，用于研究紫外线诱导的人类细胞损伤的发生率和修复情况。彗星-FISH方法能够对整个基因组以及特定染色体区域内的DNA损伤及其修复进行同步和比较分析。这种极为灵敏的方法可应用于任何损伤，比如由化学致癌物以及可转化为DNA单链或双链断裂的细胞代谢产物所诱导的损伤。文中还讨论了该方法在特定应用中的独特优势和局限性。