FOJTA, Miroslav, Luděk HAVRAN, Sabina BILLOVÁ, Pavel KOSTEČKA, Michal MASAŘÍK and René KIZEK. Two-surface strategy in electrochemical DNA hybridization assays: Detection of osmium-labeled target DNA at carbon electrodes. Electroanalysis. Germany, Weinheim: Wiley-VCH Verlag GmbH & Co., vol. 15, 5-6, p. 431-440. ISSN 1040-0397. 2003.
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Basic information
Original name Two-surface strategy in electrochemical DNA hybridization assays: Detection of osmium-labeled target DNA at carbon electrodes
Name in Czech Dvoupovrchová strategie v detekci DNA hybridizace: Detekce osmiem značených sond na uhlíkových elektrodách
Authors FOJTA, Miroslav (203 Czech Republic), Luděk HAVRAN (203 Czech Republic), Sabina BILLOVÁ (203 Czech Republic), Pavel KOSTEČKA (203 Czech Republic), Michal MASAŘÍK (203 Czech Republic, guarantor) and René KIZEK (203 Czech Republic).
Edition Electroanalysis, Germany, Weinheim, Wiley-VCH Verlag GmbH & Co. 2003, 1040-0397.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10405 Electrochemistry
Country of publisher Germany
Confidentiality degree is not subject to a state or trade secret
Impact factor Impact factor: 1.811
Organization unit Faculty of Science
UT WoS 000182694800012
Keywords in English osmium-labeled target DNA; magnetic beads; DNA hybridization; electrochemical detection; carbon electrodes
Tags carbon electrodes, DNA hybridization, electrochemical detection, magnetic beads, osmium-labeled target DNA
Tags Reviewed
Changed by Changed by: prof. RNDr. Michal Masařík, Ph.D., učo 21142. Changed: 25/6/2009 10:45.
Abstract
Target DNAs, including a 71-mer oligonucleotide, a PCR product and a plasmid DNA, all containing oligo(A) stretches,were hybridized at magnetic Dynabeads oligo(dT)(25) (DBT). The hybridization events were detected using a technique based on chemical modification of the target DNA with a complex of osmium tetroxide with 2,2'-bipyridine (Os, bipy) and voltammetric detection at carbon electrodes. DNA was modified with Os, bipy prior to capture at DBT, at the beads, or after release from the beads. In the latter case, DNA-Os, bipy was detected in the reaction mixture using adsorptive transfer stripping voltammetry involving extraction of unreacted Os, bipy from the electrode by organic solvents. Pre-labeling of the target plasmid DNA and the PCR product with Os, bipy significantly increased the yield of DNA captured at the beads. Tens of femtomoles of both short (the 71-mer oligonucleotide) and long (the 3-kilobase plasmid) target DNAs in a 20-microliter hybridization sample can be easily detected by means of these techniques: Various carbon electrode materials, including pyrolytic graphite (PGE), highly oriented pyrolytic graphite (HOPGE), carbon paste (CPE), glassy carbon and pencil graphite, were tested regarding their suitability for the detection of osmium-labeled DNA. Among them, PGE and HOPGE appeared usable in the measurements of both purified DNA-Os, bipy and its mixtures with unreacted Os, bipy while CPE was suitable for the detection purified osmium-labeled DNA.
Abstract (in Czech)
Práce popisuje elektrochemické stanovení osmiem značených oligonukleotidů a hybridizace DNA na uhlíkových elektrodách.
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