Název:
Enzymatic Incorporation of Biotin into DNA for DNA Hybridization Analysis and for Sensitive Detection of PCR-Amplified DNA
Autoři:
Špaček, Jan ; Zenka, Martin ; Haroniková, Lucia ; Havran, Luděk ; Fojta, Miroslav Typ dokumentu: Příspěvky z konference Konference/Akce: Moderní Elektrochemické Metody /34./, Jetřichovice (CZ), 2014-05-19 / 2014-05-23
Rok:
2014
Jazyk:
eng
Abstrakt: 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.
Klíčová slova:
DNA hybridization; Electrochemical DNA sensors; sensors; transferase Číslo projektu: GAP206/12/2378 (CEP), GAP206/11/1638 (CEP) Poskytovatel projektu: GA ČR, GA ČR Zdrojový dokument: XXXIV. Moderní elektrochemické metody sborník přednášek, ISBN 978-80-905221-2-1
Instituce: Biofyzikální ústav AV ČR
(web)
Informace o dostupnosti dokumentu:
Dokument je dostupný v příslušném ústavu Akademie věd ČR. Původní záznam: http://hdl.handle.net/11104/0297943