Informace o publikaci

Enzyme-linked electrochemical detection of DNA fragments amplified by PCR in the presence of a biotinylated deoxynucleoside triphosphate using disposable pencil graphite electrodes

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HÁRONÍKOVÁ Lucia ŠPAČEK Jan PLUCNARA Medard HORÁKOVÁ Petra PIVOŇKOVÁ Hana HAVRAN Luděk ERDEM Arzum FOJTA Miroslav

Rok publikování 2015
Druh Článek v odborném periodiku
Časopis / Zdroj Monatshefte fur Chemie
Fakulta / Pracoviště MU

Středoevropský technologický institut

Citace
www http://download.springer.com/static/pdf/109/art%253A10.1007%252Fs00706-015-1436-5.pdf?originUrl=http%3A%2F%2Flink.springer.com%2Farticle%2F10.1007%2Fs00706-015-1436-5&token2=exp=1451982708~acl=%2Fstatic%2Fpdf%2F109%2Fart%25253A10.1007%25252Fs00706-015-14
Doi http://dx.doi.org/10.1007/s00706-015-1436-5
Obor Biochemie
Klíčová slova Enzymes; Nucleic acids; Voltammetry
Popis In this report, we present a simple electrochemical detection protocol for the detection of specific PCR-amplified DNA fragments, based on incorporation of biotin tags into DNA amplicons during PCR run in the presence of a biotinylated nucleoside triphosphate. For detection, an enzyme-linked electrochemical system involving streptavidin-alkaline phosphatase conjugate attached to the biotinylated DNA, adsorbed at the surface of a disposable pencil graphite electrode, is used. The enzyme converts an inactive indicator, 1-naphthyl phosphate, into electrochemically oxidizable indicator 1-naphthol that is subsequently detected. Excellent selectivity of this fast, facile, and inexpensive analysis not requiring any sophisticated electrode modification and its applicability for off-line monitoring of DNA amplification is demonstrated. Applications of the technique include detection of the presence of specific nucleotide sequences in biological samples, such as sequences related to pathogenic microorganism or transgenes. [GRAPHICS] .

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