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Electrochemical Analysis of Sybr Green I and its Interaction with DNA
Dudová, Zdenka; Pivoňková, Hana; Havran, Luděk; Fojta, Miroslav
2015 - anglický
There are used many methods such real-time PCR and electrophoresis in molecular biology and biochemistry which are based on utilization of a fluorescent dye Sybr Green I (SG) for selective detection of double stranded (ds) DNA and its quantification. SG is a planar molecule (see Fig. 1) that intercalates into the DNA double helix and simultaneously can bind into minor groove of dsDNA. In our work we focused on electrochemical behavior of SG on a pyrolytic graphite electrode (PGE) using square-wave voltammetry (SWV). The voltammetric method was used in measurements of SG interactions with DNA at the PGE.
Klíčová slova:
gels; Sybr Green I; Pyrolytic graphite electrode; Square wave voltammetry
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Electrochemical Analysis of Sybr Green I and its Interaction with DNA
There are used many methods such real-time PCR and electrophoresis in molecular biology and biochemistry which are based on utilization of a fluorescent dye Sybr Green I (SG) for selective detection ...
Electrochemical Reduction and Oxidation of Nucleic Acids Bases and their Analogues: a Brief Overview
Fojta, Miroslav; Špaček, Jan; Dudová, Zdenka; Pivoňková, Hana; Daňhel, Aleš; Fojt, Lukáš; Havran, Luděk
2015 - anglický
Nucleic acids are known as electroactive biomacromolecules containing electrochemically reducible or oxidizable constituents. Nucleobases cytosine, 5-methylcytosine, adenine and guanine can be reduced in aqueous media on mercury or silver amalgam electrodes. Oxidation of all natural nucleic acids bases (in addition to the above mentioned ones, also uracil and thymine) was demonstrated using various types of carbon electrodes. Some of synthetic nucleobases or nucleotide analogues (e g., 7-deazapurines, cytidine analogues used as epigenetic modulators, etc.) exhibit specific electrochemical properties that differ from those of the parent bases and can be utilized to determine the given substance in the presence of natural nucleic acids or their components.
Klíčová slova:
elimination voltammetry; dna; oligonucleotides; structure
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Electrochemical Reduction and Oxidation of Nucleic Acids Bases and their Analogues: a Brief Overview
Nucleic acids are known as electroactive biomacromolecules containing electrochemically reducible or oxidizable constituents. Nucleobases cytosine, 5-methylcytosine, adenine and guanine can be reduced ...
Possibility of Using Purine Oxidation Signals for DNA Sequence Analysis
Špaček, Jan; Cahová, Kateřina; Havran, Luděk; Fojta, Miroslav
2015 - anglický
Signals provided by electrochemical oxidation of single stranding DNA on the surface of pyrolytic graphite electrode, which correspond to the oxidation of adenine and guanine (A and G) are known for over forty years. The mechanism of oxidation of free A and G as well as nucleoside mono-, and triphosphates of these bases is know but the mechanism of oxidation of purines in the DNA still remains unexplained. Ratiometry of A and G signals could be a new tool for DNA sequence analysis, if signals of A and G oxidation obtained from single-stranded DNA adsorbed at the electrode surface are proportional to the number of oxidized As and Gs in studied samples. Our preliminary results suggest that there is a correlation, yet we encountered problems, which yet has to be addressed and explained before this method could be applied for DNA analysis.
Klíčová slova:
DNA oxidation; DNA sequence analysis; Electrochemistry
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Possibility of Using Purine Oxidation Signals for DNA Sequence Analysis
Signals provided by electrochemical oxidation of single stranding DNA on the surface of pyrolytic graphite electrode, which correspond to the oxidation of adenine and guanine (A and G) are known for ...
Electrodeposited Coatings at ITO/FTO Enhancing their Applicability in Electroanalysis
Daňhel, Aleš; Fojta, Miroslav
2015 - anglický
Recent possibilities of modification of transparent electrodes based on indium-tin oxide and/or fluorine doped tin oxide by electrodeposited coatings enhancing their electrochemical applicability in electroanalysis are discussed in this work. Ordered mesoporous silica and/or chemically functionalized mesoporous silica thin films may serve, among the common utilization in solid phase extraction, as protective layer of the electrode surface against its fouling and/or as molecular sieve permeable for small molecules and blocking large ones (DNA, proteins, etc.). Furthermore an electrodeposition of organic modifiers and/or metal (nano) particles inside of the ordered silica nanochanels together with direct electrodeposition of selected metal (nano) particles onto selected conductive transparent supports are presented here as well.
Klíčová slova:
nucleic-acids; dna; guanine; films
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Electrodeposited Coatings at ITO/FTO Enhancing their Applicability in Electroanalysis
Recent possibilities of modification of transparent electrodes based on indium-tin oxide and/or fluorine doped tin oxide by electrodeposited coatings enhancing their electrochemical applicability in ...
Examples of Using of Electrochemical Detection at Pencil Graphite Electrode with Enzymatic Labeling for Analysis of Nucleotide Sequence
Plucnara, Medard; Haroniková, Lucia; Špaček, Jan; Havran, Luděk; Horáková, Petra; Pivoňková, Hana; Ecsin, E.; Erdem, A.; Fojta, Miroslav
2015 - anglický
Many examples of utilization of enzymatic labeling for DNA sequence analysis has been described in literature so far. Some of them involve hybridization with complementary biotinylated probe, while others use incorporation of biotinylated nucleotides into DNA strand by DNA polymerases. Common approach is then binding of streptavidine-enzyme conjugates to biotin tags and incubation with substrate, which is converted to detectable product. Here, two recent applications using this principle are described for the detection of PCR amplicons and for SNP typing. Both techniques are combined with detection at pencil graphite electrodes.
Klíčová slova:
dna hybridization; Enzymatic labeling; Alkaline phosphatase
Plné texty jsou dostupné na jednotlivých ústavech Akademie věd ČR.
Examples of Using of Electrochemical Detection at Pencil Graphite Electrode with Enzymatic Labeling for Analysis of Nucleotide Sequence
Many examples of utilization of enzymatic labeling for DNA sequence analysis has been described in literature so far. Some of them involve hybridization with complementary biotinylated probe, while ...
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