Protocol for the simultaneous quantification of oxidative purine lesions in DNA using LC-MS/MS analysis

Autor: Chryssostomos Chatgilialoglu, Marios G. Krokidis, Michael A. Terzidis
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: STAR Protocols, Vol 5, Iss 3, Pp 103191- (2024)
Druh dokumentu: article
ISSN: 2666-1667
DOI: 10.1016/j.xpro.2024.103191
Popis: Summary: Most DNA damages induced through oxidative metabolism are single lesions which can accumulate in tissues. Here, we present a protocol for the simultaneous quantification of oxidative purine lesions (cPu and 8-oxo-Pu) in DNA. We describe steps for enzymatic digestion of DNA and sample pre-purification, followed by quantification through liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. We optimized this protocol in commercially available calf thymus DNA and used genomic and mitochondrial DNA extracted from cell cultures and animal and human tissues. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
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