Quantitation of ultraviolet-induced single-strand breaks using oligonucleotide chip.

Abstract

A simple, accurate and robust methodology was established for the direct quantification of ultraviolet (UV)-induced single-strand break (SSB) using oligonucleotide chip. Oligonucleotide chips were fabricated by covalently anchoring the fluorescent-labeled ssDNAs onto silicon dioxide chip surfaces. Assuming that the possibility of more than one UV-induced SSB to be generated in a small oligonucleotide is extremely low, SSB formation was investigated quantifying the endpoint probe density by fluorescence measurement upon UV irradiation. The SSB yields obtained based on the highly sensitive laser-induced fluorometric determination of fluorophore-labeled oligonucleotides were found to coincide well with that predicted from a theoretical extrapolation of the results obtained for plasmid DNAs using conventional agarose gel electrophoresis. The developed method has the potential to serve as a high throughput, sample-thrifty, and time saving tool to realize more realistic, and direct quantification of radiation and chemical-induced strand breaks. It will be especially useful for determining the frequency of SSBs or lesions convertible to SSBs by specific cleaving reagents or enzymes.

DOI: 10.1016/j.aca.2008.05.069

Cite this paper

@article{Pal2008QuantitationOU, title={Quantitation of ultraviolet-induced single-strand breaks using oligonucleotide chip.}, author={Sukdeb Pal and Min Jung Kim and Jaebum Choo and Seong Ho Kang and Kyeong-Hee Lee and Joon Myong Song}, journal={Analytica chimica acta}, year={2008}, volume={622 1-2}, pages={195-200} }