A convenient electrogenerated chemiluminescence biosensing method for selective detection of 5-hydroxymethylcytosine in genomic DNA
Autor: | Haiqiang Deng, Meizhou Liu, Huan Wang, Wanqiao Bai, Yan Li, Huiping Sun |
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Rok vydání: | 2019 |
Předmět: |
chemistry.chemical_element
02 engineering and technology 010402 general chemistry 01 natural sciences law.invention chemistry.chemical_compound law Materials Chemistry Electrical and Electronic Engineering Instrumentation Chemiluminescence Detection limit 5-Hydroxymethylcytosine Metals and Alloys 021001 nanoscience & nanotechnology Condensed Matter Physics Combinatorial chemistry 0104 chemical sciences Surfaces Coatings and Films Electronic Optical and Magnetic Materials Ruthenium genomic DNA chemistry Electrode 0210 nano-technology Biosensor DNA |
Zdroj: | Sensors and Actuators B: Chemical. 284:236-242 |
ISSN: | 0925-4005 |
Popis: | Herein, a novel and convenient electrogenerated chemiluminescence (ECL) biosensing method for the quantitative detection of 5-hydroxymethylcytosine (5-hmC) in genomic DNA was developed by employing thiolated anchor probe DNA to immobilize 5-hydroxymethylcytosine double-stranded DNA (5-hmC-dsDNA) and a ruthenium(II) complex-tagged reporter probe as an ECL probe. Briefly, the 5-hmC-dsDNA was firstly oxidized to 5-formylcytosine dsDNA (5-fC-dsDNA) by KRuO4 in high yields. Subsequently, the thiolated anchor probe DNA is employed to help immobilize the 5-fC-dsDNA onto the gold electrode surface via the Au-S bond. An ECL probe DNA, labeled with a signal transducer − Ru(bpy)2(cbpy)NHS, hybridized with the anchor probe DNA on the electrode, resulting in an intensive ECL emission. The obtained ECL intensity was linearly proportional to the content of 5-hmC in the range from 0.0090% to 0.5761%, with the limit of detection calculated to be 0.0036% (S/N = 3). Finally, we applied the developed 5-hmC detection strategy in various mice tissues. It demonstrates that the content of 5-hmC measured by this method in different mice tissues vary significantly: 5-hmC is highest in brain and lowest in spleen. The success in assaying the mice tissues samples signifies its great promise in a broad range of practical applications for the quantitative detection of 5-hmC in genomic DNA. |
Databáze: | OpenAIRE |
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