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Understanding Signal and Background in a Thermally Resolved, Single-Branched DNA Assay Using Square Wave Voltammetry
- Source :
- Analytical Chemistry. 90:3584-3591
- Publication Year :
- 2018
- Publisher :
- American Chemical Society (ACS), 2018.
-
Abstract
- Electrochemical bioanalytical sensors with oligonucleotide transducer molecules have been recently extended for quantifying a wide range of biomolecules, from small drugs to large proteins. Short DNA or RNA strands have gained attention recently due to the existence of circulating oligonucleotides in human blood, yet challenges remain for adequately sensing these targets at electrode surfaces. In this work, we have developed a quantitative electrochemical method which uses target-induced proximity of a single-branched DNA structure to drive hybridization at an electrode surface, with readout by square-wave voltammetry (SWV). Using custom instrumentation, we first show that precise control of temperature can provide both electrochemical signal amplification and background signal depreciation in SWV readout of small oligonucleotides. Next, we thoroughly compared 25 different combinations of binding energies by their signal-to-background ratios and differences. These data served as a guide to select the optimal parameters of binding energy, SWV frequency, and assay temperature. Finally, the influence of experimental workflow on the sensitivity and limit of detection (LOD) of the sensor is demonstrated. This study highlights the importance of precisely controlling temperature and SWV frequency in DNA-driven assays on electrode surfaces while also presenting a novel instrumental design for fine-tuning of such systems.
- Subjects :
- Branched DNA Signal Amplification Assay
Oligonucleotides
010402 general chemistry
01 natural sciences
Signal
Article
Analytical Chemistry
Humans
Electrodes
Voltammetry
Detection limit
chemistry.chemical_classification
Chemistry
Oligonucleotide
business.industry
Biomolecule
010401 analytical chemistry
Temperature
Electrochemical Techniques
Equipment Design
Branched DNA assay
0104 chemical sciences
Transducer
Electrode
Optoelectronics
business
Subjects
Details
- ISSN :
- 15206882 and 00032700
- Volume :
- 90
- Database :
- OpenAIRE
- Journal :
- Analytical Chemistry
- Accession number :
- edsair.doi.dedup.....ffe1aff69153ecacd12600562d33f7ac