Original title: Enzymatic Incorporation of Biotin into DNA for DNA Hybridization Analysis and for Sensitive Detection of PCR-Amplified DNA
Authors: Špaček, Jan ; Zenka, Martin ; Haroniková, Lucia ; Havran, Luděk ; Fojta, Miroslav
Document type: Papers
Conference/Event: Moderní Elektrochemické Metody /34./, Jetřichovice (CZ), 2014-05-19 / 2014-05-23
Year: 2014
Language: eng
Abstract: We present two enzymatical electrochemical assays for DNA analysis. For hybridization analysis we used probes with biotin-14-dC introduced to 3' OH end by terminal transferase. For detection of PCR products we used Deep Vent polymerase to incorporate biotin-14-dCduring PCR. In both cases streptavidin-alkaline phosphatase conjugate was subsequently attached to the incorporated biotins and was used to catalyze dephosphorylation of 1-naphthyl phosphate to 1-naphthol, the electrochemical signal of which was utilized for detection. Compared to the former method, biotin incorporation during PCR offers lower molar detection limits, whereas application of the biotin-tailed probe can provide us with more selective detection.
Keywords: DNA hybridization; Electrochemical DNA sensors; sensors; transferase
Project no.: GAP206/12/2378 (CEP), GAP206/11/1638 (CEP)
Funding provider: GA ČR, GA ČR
Host item entry: XXXIV. Moderní elektrochemické metody sborník přednášek, ISBN 978-80-905221-2-1

Institution: Institute of Biophysics AS ČR (web)
Document availability information: Fulltext is available at the institute of the Academy of Sciences.
Original record: http://hdl.handle.net/11104/0297943

Permalink: http://www.nusl.cz/ntk/nusl-396442


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Research > Institutes ASCR > Institute of Biophysics
Conference materials > Papers