Fluorometric determination of nucleic acids based on the use of polydopamine nanotubes and target-induced strand displacement amplification
| Autor: | Jia Ge, Xin Geng, Lin Zhang, Dong-Mei Bai, Yalei Hu, Zhaohui Li, Ke Xing, Qi-Yong Cai |
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| Rok vydání: | 2017 |
| Předmět: |
Nucleic acid quantitation
Indoles Polymers DNA Single-Stranded 02 engineering and technology Diamines 010402 general chemistry 01 natural sciences Analytical Chemistry Polymerization chemistry.chemical_compound Molecular beacon Nucleic Acids Fluorometry Benzothiazoles Organic Chemicals Polymerase Klenow fragment Nanotubes biology Hybridization probe Multiple displacement amplification 021001 nanoscience & nanotechnology 0104 chemical sciences chemistry SYBR Green I biology.protein Biophysics Quinolines Primer (molecular biology) 0210 nano-technology DNA Probes Nucleic Acid Amplification Techniques |
| Zdroj: | Mikrochimica acta. 185(2) |
| ISSN: | 1436-5073 |
| Popis: | The authors describe a fluorometric method for the quantitation of nucleic acids by combining (a) cycled strand displacement amplification, (b) the unique features of the DNA probe SYBR Green, and (c) polydopamine nanotubes. SYBR Green undergoes strong fluorescence enhancement upon intercalation into double-stranded DNA (dsDNA). The polydopamine nanotubes selectively adsorb single-stranded DNA (ssDNA) and molecular beacons. In the absence of target DNA, the molecular beacon, primer and SYBR Green are adsorbed on the surface of polydopamine nanotubes. This results in quenching of the fluorescence of SYBR Green, typically measured at excitation/emission wavelengths of 488/518 nm. Upon addition of analyte (target DNA) and polymerase, the stem of the molecular beacon is opened so that it can bind to the primer. This triggers target strand displacement polymerization, during which dsDNA is synthesized. The hybridized target is then displaced due to the strand displacement activity of the polymerase. The displaced target hybridizes with another molecular beacon. This triggers the next round of polymerization. Consequently, a large amount of dsDNA is formed which is detected by addition of SYBR Green. Thus, sensitive and selective fluorometric detection is realized. The fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 0.05 to 25 nM with a low limit of detection of 20 pM. Graphical abstract Schematic of a fluorometric strategy for highly sensitive and selective determination of nucleic acids by combining strand displacement amplification and the unique features of SYBR Green I (SG) and polydopamine nanotubes. |
| Databáze: | OpenAIRE |
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