Rapid Quantification of Oxidation and UV Induced DNA Damage by Repair Assisted Damage Detection-(Rapid RADD)

Noa Gilat, Dmitry Torchinsky, Sapir Margalit, Yael Michaeli, Sigal Avraham, Hila Sharim, Nadav Elkoshi, Carmit Levy, Shahar Zirkin, Yuval Ebenstein

Research output: Contribution to journalArticlepeer-review

Abstract

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.

Original languageEnglish
Pages (from-to)9887-9894
Number of pages8
JournalAnalytical Chemistry
Volume92
Issue number14
DOIs
StatePublished - 21 Jul 2020

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