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Title: Oxidative and nitrosative DNA damage: Occurrence, measurement and mechanism
Keywords: Mitochondrial DNA, oxidative damage, free radical, DNA deamination, GC/MS, artifacts
Issue Date: 13-Feb-2006
Citation: LIM KOK SEONG (2006-02-13). Oxidative and nitrosative DNA damage: Occurrence, measurement and mechanism. ScholarBank@NUS Repository.
Abstract: (1) Mitochondrial DNA (mtDNA) has been believed to suffer greater endogenous oxidative damage than nuclear DNA (nDNA) but the large variation among the published damage levels has cast doubt on the validity of the methods used. Using mtDNA of high purity, we show that mtDNA damage is not higher than nDNA damage. (2) Ex vivo artefactual oxidation of mtDNA during isolation of mitochondria has been suggested to explain the observed high levels of damage in mtDNA. Our study shows that artefactual oxidation of biomolecules does not occur and generation of hydrogen peroxide was not detectable during the isolation of rat liver mitochondria. (3) Acid hydrolysis of DNA is commonly used to release DNA bases for GC/MS analysis but artefactual DNA deamination has rarely been studied. We demonstrate the occurrence of artefactual DNA deamination caused by hot formic acid and developed an improved GC/MS method to measure 2a??-deoxyinosine.
Appears in Collections:Ph.D Theses (Open)

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