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Title of article :
Quantitation of ultraviolet-induced single-strand breaks using oligonucleotide chip Original Research Article
Author/Authors :
Sukdeb Pal، نويسنده , , Min Jung Kim، نويسنده , , Jaebum Choo، نويسنده , , Seong Ho Kang، نويسنده , , Kyeong-Hee Lee، نويسنده , , Joon Myong Song، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2008
Pages :
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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.
Keywords :
Surface coverage , Alkaline phosphatase-digestion , Single-strand breaks , Oligonucleotide chip , Laser-induced fluorescence
Journal title :
Analytica Chimica Acta
Link To Document :