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Nucleic Acids Research, 2001, Vol. 29, No. 10 2117-2126
© 2001 Oxford University Press

A reliable assessment of 8-oxo-2-deoxyguanosine levels in nuclear and mitochondrial DNA using the sodium iodide method to isolate DNA

Michelle L. Hamilton1, ZhongMao Guo1,4, Clinton D. Fuller1, Holly Van Remmen1,4, Walter F. Ward1, Steven N. Austad5, Dean A. Troyer2,3, Ian Thompson3 and Arlan Richardson1,4,*

1Department of Physiology, 2Department of Pathology and 3Division of Urology, Department of Surgery, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USA, 4South Texas Veterans Health Care System, San Antonio, TX 78284-7762, USA and 5Department of Biological Sciences, University of Idaho, Moscow, ID 83844, USA

A major controversy in the area of DNA biochemistry concerns the actual in vivo levels of oxidative damage in DNA. We show here that 8-oxo-2-deoxyguanosine (oxo8dG) generation during DNA isolation is eliminated using the sodium iodide (NaI) isolation method and that the level of oxo8dG in nuclear DNA (nDNA) is almost one-hundredth of the level obtained using the classical phenol method. We found using NaI that the ratio of oxo8dG/105 deoxyguanosine (dG) in nDNA isolated from mouse tissues ranged from 0.032 ± 0.002 for liver to 0.015 ± 0.003 for brain. We observed a significant increase (10-fold) in oxo8dG in nDNA isolated from liver tissue after 2 Gy of {gamma}-irradiation when NaI was used to isolate DNA. The turnover of oxo8dG in nDNA was rapid, e.g. disappearance of oxo8dG in the mouse liver in vivo after {gamma}-irradiation had a half-life of 11 min. The levels of oxo8dG in mitochondrial DNA isolated from liver, heart and brain were 6-, 16- and 23-fold higher than nDNA from these tissues. Thus, our results showed that the steady-state levels of oxo8dG in mouse tissues range from 180 to 360 lesions in the nuclear genome and from one to two lesions in 100 mitochondrial genomes.

* To whom correspondence should be addressed at: Department of Physiology, MSC-7756, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78229-3900, USA. Tel: +1 210 567 4397; Fax: +1 210 567 4414; Email: richardsona{at}uthscsa.edu


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