Quantification of UV-induced cyclobutane pyrimidine dimers using an oligonucleotide chip assay
- Authors
- Kim, Min Jung; Lee, Su Chul; Kang, Seong Ho; Choo, Jaebum; Song, Joon Myong
- Issue Date
- Jul-2010
- Publisher
- SPRINGER HEIDELBERG
- Keywords
- Oligonucleotide chip; Lesion-specific enzyme-induced break assay; Cyclobutane pyrimidine dimer; T4 endonuclease V
- Citation
- ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v.397, no.6, pp 2271 - 2277
- Pages
- 7
- Journal Title
- ANALYTICAL AND BIOANALYTICAL CHEMISTRY
- Volume
- 397
- Number
- 6
- Start Page
- 2271
- End Page
- 2277
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/46857
- DOI
- 10.1007/s00216-010-3793-6
- ISSN
- 1618-2642
1618-2650
- Abstract
- A lesion-specific enzyme-induced DNA strand break assay was developed for an oligonucleotide chip for the determination of UVB-induced cyclobutane pyrimidine dimers (CPDs). A 20-mer of fluorophore-labeled and biotinylated oligonucleotide was immobilized on the chip. CPDs in DNA on the chip were formed by UVB irradiation (312 nm). T4 endonuclease V (T4N5) was used to excise the CPD site as T4N5 sensitively and specifically detects CPDs. The fluorophore-labeled DNA fragments were detected by a laser-induced fluorescence (LIF) detection system. The number of CPDs induced by UVB was determined based on a mathematical equation obtained from a predetermined calibration curve. The yield of UVB-induced CPDs was 1.73 CPDs per megabase per (kJ/m(2)). The reliability of this value was proved by its similarity to reference values obtained from gel electrophoresis. The developed assay has strong potential to quantify most kinds of UV-induced DNA lesions.
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