Knowing the amount and type of DNA damage is of great significance for a broad range of clinical and research applications. However, existing methods are either lacking in their ability to distinguish between types of DNA damage or limited in their sensitivity and reproducibility. The method described herein enables rapid and robust quantification of type-specific single-strand DNA damage. The method is based on repair-assisted damage detection (RADD) by which fluorescent nucleotides are incorporated into DNA damage sites using type-specific repair enzymes. Up to 90 DNA samples are then deposited on a multiwell glass slide, and analyzed by a conventional slide scanner for quantification of DNA damage levels. Accurate and sensitive measurements of oxidative or UV-induced DNA damage levels and repair kinetics are presented for both in vitro and in vivo models.

中文翻译：

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通过修复辅助损伤检测（快速RADD）快速定量氧化和紫外线诱导的DNA损伤。

了解DNA损伤的程度和类型对于广泛的临床和研究应用具有重要意义。但是，现有方法要么缺乏区分DNA损伤类型的能力，要么缺乏灵敏度和可重复性。本文所述的方法能够对类型特异性单链DNA损伤进行快速而稳健的定量。该方法基于修复辅助损伤检测（RADD），通过该检测可以使用类型特异性修复酶将荧光核苷酸掺入DNA损伤位点。然后将多达90个DNA样品沉积在多孔玻璃载玻片上，并通过常规载玻片扫描仪进行分析以定量DNA损伤水平。