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National Repository of Grey Literature 3 records found  Search took 0.01 seconds. 
Enzymatic Incorporation of Biotin into DNA for DNA Hybridization Analysis and for Sensitive Detection of PCR-Amplified DNA
 Špaček, Jan ; Zenka, Martin ; Haroniková, Lucia ; Havran, Luděk ; Fojta, Miroslav
We present two enzymatical electrochemical assays for DNA analysis. For hybridization analysis we used probes with biotin-14-dC introduced to 3' OH end by terminal transferase. For detection of PCR products we used Deep Vent polymerase to incorporate biotin-14-dCduring PCR. In both cases streptavidin-alkaline phosphatase conjugate was subsequently attached to the incorporated biotins and was used to catalyze dephosphorylation of 1-naphthyl phosphate to 1-naphthol, the electrochemical signal of which was utilized for detection. Compared to the former method, biotin incorporation during PCR offers lower molar detection limits, whereas application of the biotin-tailed probe can provide us with more selective detection.
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Voltammetric Analysis of Anthraquinone-labeled Nucleotide Triphosphates and Oligonucleotides at Gold Electrodes
 Vidláková, Pavlína ; Balintová, Jana ; Havran, Luděk ; Hocek, Michal ; Fojta, Miroslav
DNA labelling is used for increase of sensitivity of electrochemical detection. When the DNA or oligonucleotides (ONs) are chemically modified, their electrochemical responses can be changed depending on electrochemical activity of the introduced moiety. One option of DNA labeling is incorporation of chemically modified nucleotides using corresponding deoxynucleotide triphosphates (dNTPs) into DNA molecules with using primer extension (PEX). In our study we used anthraquinone labeled dNTPs and ONs with enzymatically incorporated anthraquinone labeled nucleotides. Thus synthesized ON stretch bearing the anthraquinone tags was electrochemically analyzed using voltammetry at gold electrodes.
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Redox DNA Labeling - from Simple DNA Detection by Osmium Tetroxide Complexes Modification to Ratiometric Sequence Analysis
 Havran, Luděk ; Balintová, Jana ; Vidláková, Pavlína ; Macíčková-Cahová, Hana ; Pivoňková, Hana ; Hocek, Michal ; Fojta, Miroslav
DNA is electroactive molecule producing analytically useful intrinsic voltammetric signals. For some applications is a useful use redox active tag to improve specificity of the analysis. One from approaches how to prepare DNA bearing redox label is its modification by complexes of osmium tetroxide with nitrogen ligands. This method find wide used in highly sensitive DNA detection and in development of electrochemical DNA hybridization sensors. Another possibility for preparation of redox labeled DNA is incorporation of redox tags modified deoxynucleotide triphosphates by DNA polymerases. In this contribution will be demonstrated application of various redox labels in electrochemical analysis of DNA.
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