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

使用 LC-MS/MS 分析同时定量 DNA 中的氧化嘌呤损伤的方案

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作者:Chryssostomos Chatgilialoglu, Marios G Krokidis, Michael A Terzidis

Abstract

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.

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