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Enzyme-linked electrochemical DNA ligation assay using magnetic beads
- 1.0431757 - BFÚ 2015 RIV DE eng J - Journal Article
Stejskalová, Eva - Horáková Brázdilová, Petra - Vacek, J. - Bowater, R. P. - Fojta, Miroslav
Enzyme-linked electrochemical DNA ligation assay using magnetic beads.
Analytical and Bioanalytical Chemistry. Roč. 406, č. 17 (2014), s. 4129-4136. ISSN 1618-2642. E-ISSN 1618-2650
R&D Projects: GA ČR(CZ) GPP206/11/P739; GA ČR(CZ) GAP206/11/1638; GA AV ČR(CZ) IAA400040901
Institutional support: RVO:68081707
Keywords : Electrochemistry * Enzyme labeling * DNA ligase
Subject RIV: BO - Biophysics
Impact factor: 3.436, year: 2014
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.
Permanent Link: http://hdl.handle.net/11104/0236329
Number of the records: 1