Open AccessA qPCR-based assay to quantify oxidized guanine and other FPG-sensitive base lesions within telomeric DNA
Author(s) -
Nathan O’Callaghan,
Natalie Baack,
Razinah Sharif,
Michael Fenech
Publication year - 2011
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000113788
Subject(s) - telomere , dna glycosylase , dna , dna damage , guanine , oxidative stress , microbiology and biotechnology , biology , biochemistry , hydrogen peroxide , chemistry , gene , nucleotide
Telomere shortening is an important risk factor for cancer and accelerated aging. However, it is becoming evident that oxidatively damaged DNA within the telomere sequence may also cause telomere dysfunction. Here we describe a reliable, cost-effective quantitative PCR (qPCR)-based method to measure the amount of oxidized residues within telomeric DNA that are recognized and excised by formamidopyridine DNA glycosylase (FPG). We also report that in an in vitro model of oxidative stress oxidized base lesions measured using this method are more prevalent within telomeric sequences. Furthermore, this method is sufficiently sensitive to detect changes in oxidative stress induced by zinc deficiency and hydrogen peroxide within the physiological range.