Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.freeradbiomed.2006.01.030
Title: Potential artifacts in the measurement of DNA deamination
Authors: Lim, K.S.
Huang, S.H. 
Jenner, A. 
Wang, H. 
Tang, S.Y. 
Halliwell, B. 
Keywords: 2′-Deoxyinosine
DNA
Free radicals
GC-MS
LPS
Issue Date: 2006
Source: Lim, K.S.,Huang, S.H.,Jenner, A.,Wang, H.,Tang, S.Y.,Halliwell, B. (2006). Potential artifacts in the measurement of DNA deamination. Free Radical Biology and Medicine 40 (11) : 1939-1948. ScholarBank@NUS Repository. https://doi.org/10.1016/j.freeradbiomed.2006.01.030
Abstract: Attack on DNA by some reactive nitrogen species results in deamination of adenine and guanine, leading to the formation of hypoxanthine and xanthine, respectively. Published levels of these products in cellular DNA have varied widely. Although these two deamination products are often measured by GC-MS analysis, the procedure of acid hydrolysis to release DNA bases for derivatization poses a risk of artifactual deamination of the DNA. In this study, we demonstrated the artifactual formation of these two deamination products during acid hydrolysis and hence developed a method for detecting and measuring 2′-deoxyinosine, the nucleoside of hypoxanthine. Our assay for 2′-deoxyinosine employs nuclease P1 and alkaline phosphatase to achieve release of the nucleosides from DNA, followed by HPLC prepurification with subsequent GC-MS analysis of the nucleosides. This assay detected an increase in the levels of 2′-deoxyinosine in DNA when commercial salmon testis DNA was treated with nitrous acid. We also used it to measure levels in various rat tissues of both normal and endotoxin-treated rats, but could not find increased 2′-deoxyinosine formation in tissues even though ·NO production was substantially increased. © 2006 Elsevier Inc. All rights reserved.
Source Title: Free Radical Biology and Medicine
URI: http://scholarbank.nus.edu.sg/handle/10635/28638
ISSN: 08915849
DOI: 10.1016/j.freeradbiomed.2006.01.030
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.

SCOPUSTM   
Citations

20
checked on Dec 13, 2017

WEB OF SCIENCETM
Citations

18
checked on Nov 5, 2017

Page view(s)

163
checked on Dec 9, 2017

Google ScholarTM

Check

Altmetric


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.