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Statements

Subject Item
n2:RIV%2F00216224%3A14740%2F14%3A00075856%21RIV15-MSM-14740___
rdf:type
skos:Concept n9:Vysledek
rdfs:seeAlso
http://link.springer.com/article/10.1007%2Fs00216-014-7811-y
dcterms: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.
dcterms:title
Enzyme-linked electrochemical DNA ligation assay using magnetic beads Enzyme-linked electrochemical DNA ligation assay using magnetic beads
skos:prefLabel
Enzyme-linked electrochemical DNA ligation assay using magnetic beads Enzyme-linked electrochemical DNA ligation assay using magnetic beads
skos:notation
RIV/00216224:14740/14:00075856!RIV15-MSM-14740___
n4:aktivita
n18:P n18:I
n4:aktivity
I, P(GAP206/11/1638), P(GPP206/11/P739), P(IAA400040901)
n4:cisloPeriodika
17
n4:dodaniDat
n10:2015
n4:domaciTvurceVysledku
n7:4171373
n4:druhVysledku
n20:J
n4:duvernostUdaju
n11:S
n4:entitaPredkladatele
n16:predkladatel
n4:idSjednocenehoVysledku
14820
n4:idVysledku
RIV/00216224:14740/14:00075856
n4:jazykVysledku
n19:eng
n4:klicovaSlova
Electrochemistry; Enzyme labeling; DNA ligase; Nicked DNA; Magnetic beads; Immobilized assays
n4:klicoveSlovo
n13:Enzyme%20labeling n13:Electrochemistry n13:Nicked%20DNA n13:Immobilized%20assays n13:DNA%20ligase n13:Magnetic%20beads
n4:kodStatuVydavatele
DE - Spolková republika Německo
n4:kontrolniKodProRIV
[CDBE778CE7FC]
n4:nazevZdroje
Analytical and Bioanalytical Chemistry
n4:obor
n6:CB
n4:pocetDomacichTvurcuVysledku
1
n4:pocetTvurcuVysledku
5
n4:projekt
n5:IAA400040901 n5:GAP206%2F11%2F1638 n5:GPP206%2F11%2FP739
n4:rokUplatneniVysledku
n10:2014
n4:svazekPeriodika
406
n4:tvurceVysledku
Bowater, Richard Horáková, Petra Vacek, Jan Fojta, Miroslav Stejskalová, Eva
n4:wos
000337787400010
s:issn
1618-2642
s:numberOfPages
8
n12:doi
10.1007/s00216-014-7811-y
n8:organizacniJednotka
14740