Use of a capacitive affinity biosensor for sensitive and selective detection and quantification of DNA-A model study
| Autor: | Eva Ståhl-Wernersson, Martin Hedström, Ally Mahadhy, Bo Mattiasson |
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| Rok vydání: | 2014 |
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| Zdroj: | Biotechnology Reports Biotechnology Reports, Vol 3, Iss C, Pp 42-48 (2014) |
| ISSN: | 2215-017X |
| Popis: | Highlights • A DNA-sensor was developed to monitor the hybridization of complementary ssDNAs. • The signal strength was proportional to the length of short oligonucleotides. • Elevated temperature was observed to reduce non-specific interactions. • The study proposes sandwich hybridization approach to improve signal strength. A capacitive DNA-sensor model system was used to monitor the capture of complementary single-stranded DNAs. The sensor chip consisted of a gold electrode, which was carefully insulated with a polytyramine layer and covalently tagged with 25-mer oligo-C. As low as 10−11 moles per liter of target oligo-G could be detected by injecting 250 μL of sample. Elevated temperature was used to reduce non-specific hybridization. Less than 10% of non-target 25-mer oligo-T interacted nonspecifically with the oligo-C probes when hybridization process was performed at 50 °C. Studying the relationship of length of the analyte to the signal strength, the output from the capacitive DNA-sensor increased to almost the double; from 50 to 88-nF cm−2, when a 25-mer oligo-G was used instead of a 15-mer. By sandwich hybridization at room temperature, it was possible to further increase the signal, from 78-nF cm−2 for the target 50-mer oligo-G alone, to 114-nF cm−2. |
| Databáze: | OpenAIRE |
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