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Description
| - DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5 biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3 digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode.
- DNA ligases are essential enzymes in all cells and have been proposed as targets for novel antibiotics. Efficient DNA ligase activity assays are thus required for applications in biomedical research. Here we present an enzyme-linked electrochemical assay based on two terminally tagged probes forming a nicked junction upon hybridization with a template DNA. Nicked DNA bearing a 5 biotin tag is immobilized on the surface of streptavidin-coated magnetic beads, and ligated product is detected via a 3 digoxigenin tag recognized by monoclonal antibody-alkaline phosphatase conjugate. Enzymatic conversion of napht-1-yl phosphate to napht-1-ol enables sensitive detection of the voltammetric signal on a pyrolytic graphite electrode. (en)
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Title
| - Enzyme-linked electrochemical DNA ligation assay using magnetic beads
- Enzyme-linked electrochemical DNA ligation assay using magnetic beads (en)
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skos:prefLabel
| - Enzyme-linked electrochemical DNA ligation assay using magnetic beads
- Enzyme-linked electrochemical DNA ligation assay using magnetic beads (en)
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skos:notation
| - RIV/00216224:14740/14:00075856!RIV15-MSM-14740___
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http://linked.open...avai/riv/aktivita
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http://linked.open...avai/riv/aktivity
| - I, P(GAP206/11/1638), P(GPP206/11/P739), P(IAA400040901)
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http://linked.open...iv/cisloPeriodika
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http://linked.open...vai/riv/dodaniDat
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http://linked.open...aciTvurceVysledku
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http://linked.open.../riv/druhVysledku
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http://linked.open...iv/duvernostUdaju
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http://linked.open...titaPredkladatele
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http://linked.open...dnocenehoVysledku
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http://linked.open...ai/riv/idVysledku
| - RIV/00216224:14740/14:00075856
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http://linked.open...riv/jazykVysledku
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http://linked.open.../riv/klicovaSlova
| - Electrochemistry; Enzyme labeling; DNA ligase; Nicked DNA; Magnetic beads; Immobilized assays (en)
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http://linked.open.../riv/klicoveSlovo
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http://linked.open...odStatuVydavatele
| - DE - Spolková republika Německo
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http://linked.open...ontrolniKodProRIV
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http://linked.open...i/riv/nazevZdroje
| - Analytical and Bioanalytical Chemistry
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http://linked.open...in/vavai/riv/obor
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http://linked.open...ichTvurcuVysledku
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http://linked.open...cetTvurcuVysledku
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http://linked.open...vavai/riv/projekt
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http://linked.open...UplatneniVysledku
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http://linked.open...v/svazekPeriodika
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http://linked.open...iv/tvurceVysledku
| - Fojta, Miroslav
- Horáková, Petra
- Stejskalová, Eva
- Vacek, Jan
- Bowater, Richard
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http://linked.open...ain/vavai/riv/wos
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issn
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number of pages
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http://bibframe.org/vocab/doi
| - 10.1007/s00216-014-7811-y
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http://localhost/t...ganizacniJednotka
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