TY - JOUR
T1 - Rapid Quantification of Oxidation and UV Induced DNA Damage by Repair Assisted Damage Detection-(Rapid RADD)
AU - Gilat, Noa
AU - Torchinsky, Dmitry
AU - Margalit, Sapir
AU - Michaeli, Yael
AU - Avraham, Sigal
AU - Sharim, Hila
AU - Elkoshi, Nadav
AU - Levy, Carmit
AU - Zirkin, Shahar
AU - Ebenstein, Yuval
N1 - Publisher Copyright:
© 2020 American Chemical Society.
PY - 2020/7/21
Y1 - 2020/7/21
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85089813701&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.0c01393
DO - 10.1021/acs.analchem.0c01393
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C2 - 32578422
AN - SCOPUS:85089813701
SN - 0003-2700
VL - 92
SP - 9887
EP - 9894
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 14
ER -