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Proceedings Paper

Optical detection of DNA damage
Author(s): Kim R. Rogers; A. Apostol; J. Cembrano
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Paper Abstract

A rapid and sensitive fluorescence assay for oxidative damage to calf thymus DNA is reported. A decrease in the transition temperature for strand separation resulted from exposure of the DNA to the reactive decomposition products of 3- morpholinosydnonimine (SIN-1) (i.e., nitric oxide, superoxide, peroxynitrite, hydrogen peroxide, and hydroxyl radicals). A decrease in melting temperature of 12 degrees Celsius was indicative of oxidative damage including single strand chain breaks. Double stranded (ds) and single stranded (ss) forms of DNA were determined using the indicator dyes ethidium bromide and PicoGreen. The change in DNA 'melting' curves was dependant on the concentration of SIN-1 and was most pronounced at 75 degrees Celsius. This chemically induced damage was significantly inhibited by sodium citrate, tris(hydroxymethyl)aminomethane (Tris), and diethylenetriaminepentaacetic acid (DTPA), but was unaffected by superoxide dismutase (SOD), catalase, ethylenediamine tetraacietic acid (EDTA), or deferoxamine. Lowest observable effect level for SIN-1-induced damage was 200 (mu) M.

Paper Details

Date Published: 10 February 1999
PDF: 5 pages
Proc. SPIE 3534, Environmental Monitoring and Remediation Technologies, (10 February 1999); doi: 10.1117/12.338987
Show Author Affiliations
Kim R. Rogers, U.S. Environmental Protection Agency (United States)
A. Apostol, U.S. Environmental Protection Agency (United States)
J. Cembrano, U.S. Environmental Protection Agency (United States)

Published in SPIE Proceedings Vol. 3534:
Environmental Monitoring and Remediation Technologies
Tuan Vo-Dinh; Robert L. Spellicy, Editor(s)

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