National Repository of Grey Literature 2 records found  Search took 0.00 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.
New Ligands for Modification of DNA by Osmium Tetroxide Reagents
Havran, Luděk ; Zenka, Martin ; Fojta, Miroslav
Besides DNA intrinsic electroactivity, modification of DNA by redox active tags find wide use in electrochemical analysis of DNA interactions and damage. Application of these tags can improve specificity of the analysis. One from utilized labels in this field are complexes of osmium tetroxide with nitrogen ligands, which produce with DNA electroactive covalent adducts. These complexes preferentially react with thymine in single strand DNA. Type of used ligand influence electrochemical behavior forming adducts. In this contribution will be presented results which are acquired by testing of set a new ligands based of quinone and substituted 1, 10-phenanthroline.

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