Bilichak Andriy, Kovalchuk Igor
Lethbridge Research Centre, Agriculture and Agri-Food Canada, Lethbridge, AB, Canada.
Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, AB, Canada, T1K 3M4.
Methods Mol Biol. 2017;1456:237-242. doi: 10.1007/978-1-4899-7708-3_18.
DNA strand breaks arise from normal cellular processes such as replication, transcription, and DNA repair as well as spontaneous DNA damage caused by cell metabolic activities. In addition, strand breaks occur due to direct or indirect DNA damage produced by various abiotic and biotic stresses. Strand breaks are among the most problematic DNA lesions because unrepaired strand breaks may lead to cell cycle arrest, gross chromosome rearrangements, or even cell death. Thus, the measurement of the relative number of strand breaks can provide an informative picture of genome stability of a given cell, tissue, or organism. Here, we describe the use of random oligonucleotide-primed synthesis (ROPS) assay for the detection and quantification of the level of strand breaks in tissue samples. The applications of the assay for a quantitative detection of 3'OH, 3'P, or DNA strand breaks at a cleavage site of the deoxyribose residue are discussed.
DNA链断裂源于正常的细胞过程，如复制、转录和DNA修复，以及细胞代谢活动引起的自发DNA损伤。此外，各种非生物和生物胁迫产生的直接或间接DNA损伤也会导致链断裂。链断裂是最成问题的DNA损伤之一，因为未修复的链断裂可能导致细胞周期停滞、染色体大规模重排，甚至细胞死亡。因此，测量链断裂的相对数量可以提供给定细胞、组织或生物体基因组稳定性的信息。在这里，我们描述了使用随机寡核苷酸引物合成（ROPS）分析来检测和定量组织样本中链断裂的水平。讨论了该分析在定量检测脱氧核糖残基切割位点处的3'OH、3'P或DNA链断裂方面的应用。
