title: A label-free fluorescence strategy for selective detection of nicotinamide adenine dinucleotide based on a dumbbell-like probe with low background noise.
abstract: In this work we developed a novel label-free fluorescence sensing approach for the detection of nicotinamide adenine dinucleotide (NAD(+)) based on a dumbbell-like DNA probe designed for both ligation reaction and digestion reaction with low background noise. SYBR Green I (SG I), a double-helix dye, was chosen as the readout fluorescence signal. In the absence of NAD(+), the ligation reaction did not occur, but the probe was digested to mononucleotides after the addition of exonuclease I (Exo I) and exonuclease I (Exo III), resulting in a weak fluorescence intensity due to the weak interaction between SG I and mononucleotides. In the presence of NAD(+), the DNA probe was ligated by Escherichia coli DNA ligase, blocking the digestion by Exo I and Exo III. As a result, SG I was intercalated into the stem part of the DNA dumbbell probe and fluorescence enhancement was achieved. This method was simple in design, fast to operate, with good sensitivity and selectivity which could discriminate NAD(+) from its analogs.
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Xuexu
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China; School of Physics and Centre for Climate and Air Pollution Studies, Ryan Institute, National University of Ireland Galway, University Road, Galway, Ireland.
authors.forename: Chunshui
authors.initials: C
authors.lastname: Lin
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
authors.forename: Yiying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China. Electronic address: yrwang@xmu.edu.cn.
authors.forename: Yiru
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Chemistry and the MOE Key Laboratory of Spectrochemical Analysis & Instrumentation, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China; State Key Laboratory of Marine Environmental Science, Xiamen University, Xiamen 361005, China. Electronic address: xichen@xmu.edu.cn.
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
mesh_terms: D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics / Q000648:ultrastructure*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D009243:NAD / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D059629:Signal-To-Noise Ratio; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*; D013194:Staining and Labeling
chemical_list: D015342:DNA Probes; D009243:NAD
keywords: Fluorescence; Label-free; Nicotinamide adenine dinucleotide (NAD(+)); SYBR Green I