No (298)
Number of found documents:
298
Published from
to
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
2014 - English
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.
Keywords:
transferase; sensors; DNA hybridization; Electrochemical DNA sensors
Available at various institutes of the ASCR
Enzymatic Incorporation of Biotin into DNA for DNA Hybridization Analysis and for Sensitive Detection of PCR-Amplified DNA
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 ...
Image Analysis of Gene Locus Positions Within Chromosome Territories in Human Lymphocytes
Stepka, K.; Falk, Martin
2014 - English
One of the important areas of current cellular research with substantial impacts on medicine is analyzing the spatial organization of genetic material within the cell nuclei. Higher-order chromatin structure has been shown to play essential roles in regulating fundamental cellular processes, like DNA transcription, replication, and repair. In this paper, we present an image analysis method for the localization of gene loci with regard to chromosomal territories they occupy in 3D confocal microscopy images. We show that the segmentation of the territories to obtain a precise position of the gene relative to a hard territory boundary may lead to undesirable bias in the results, instead, we propose an approach based on the evaluation of the relative chromatin density at the site of the gene loci. This method yields softer, fuzzier ´boundaries´, characterized by progressively decreasing chromatin density. The method therefore focuses on the extent to which the signals are located inside the territories, rather than a hard yes/no classification.
Keywords:
selection method; deletion
Available at various institutes of the ASCR
Image Analysis of Gene Locus Positions Within Chromosome Territories in Human Lymphocytes
One of the important areas of current cellular research with substantial impacts on medicine is analyzing the spatial organization of genetic material within the cell nuclei. Higher-order chromatin ...
P36 VOLTAMETRIC DETECTION OF DNA DELETION ON PENCIL ELECTRODE
Haroniková, Lucia; Špaček, Jan; Fojta, Miroslav
2014 - English
In this work, we present a new qualitative approach of detection of PCR products using electrochemistry on pencil electrodes. PCR products with an incorporated biotin-labeled dNTP (dCTP in this study) are detected via conjugated streptavidinealkaline phospatase. 1-Naphthyl phosphate (1-NP), which is dephosphorylated by alkaline phosphatase to release 1-naphthol, was used as a substrate in electrochemical detection of the PCR product. Voltammetric measurement on disposable pencil electrode is a very cheap and easy to use method. The system was optimized for plasmid DNA PCR product with potential to detect human genome DNA products and possible application in gene deletion monitoring.
Keywords:
DNA
Available at various institutes of the ASCR
P36 VOLTAMETRIC DETECTION OF DNA DELETION ON PENCIL ELECTRODE
In this work, we present a new qualitative approach of detection of PCR products using electrochemistry on pencil electrodes. PCR products with an incorporated biotin-labeled dNTP (dCTP in this study) ...
Detection of Single Nucleotide Polymorphisms Using Selective Incorporation of Biotin in DNA Strand and Subsequent Enzymatic Detection at Pencil Electrode
Plucnara, Medard; Ecsin, E.; Erdem, A.; Fojta, Miroslav
2014 - English
Enzyme-linked electrochemical assay using DNA labeling by biotin followed by binding of enzyme-streptavidin conjugates has been found to be a potent tool for DNA diagnostic. This approach brings a special advantage of signal amplification due to the fact, that only very small number of enzymatic labels can produce a number of molecules of an electrochemically detectable product from an inactive substrate to obtain sufficiently strong signal. A new way of using of this technique combined with selective primer extension reaction designed for the detection of single nucleotide polymorphism is presented here. The assay was combined with measurements at a pencil graphite electrode, which is a very practical tool for potential clinical applications due to its cheapness and disposability.
Keywords:
Single nucleotide polymorphism; Enzymatic labeling; Pencil graphite electrode
Available at various institutes of the ASCR
Detection of Single Nucleotide Polymorphisms Using Selective Incorporation of Biotin in DNA Strand and Subsequent Enzymatic Detection at Pencil Electrode
Enzyme-linked electrochemical assay using DNA labeling by biotin followed by binding of enzyme-streptavidin conjugates has been found to be a potent tool for DNA diagnostic. This approach brings a ...
TAIL-LABELED OLIGONUCLEOTIDE PROBES FOR A DUAL ELECTROCHEMICAL MAGNETIC IMMUNOPRECIPITATION ASSAY OF DNA-PROTEIN BINDING
Hermanová, Monika; Špaček, Jan; Orság, Petr; Fojta, Miroslav
2014 - English
A novel assay for detection of DNA-protein binding has been developed. Oligonucleotides bearing or lacking specific binding site of the p53 protein were tail-labeled by two different modified deoxynucleotide triphosphates using terminal deoxynucleotidyl transferase. Electrochemical detection enabled to discriminate between sequence-specific and non-specific p53-DNA binding in a competition assay.
Keywords:
DNA-PROTEIN
Available at various institutes of the ASCR
TAIL-LABELED OLIGONUCLEOTIDE PROBES FOR A DUAL ELECTROCHEMICAL MAGNETIC IMMUNOPRECIPITATION ASSAY OF DNA-PROTEIN BINDING
A novel assay for detection of DNA-protein binding has been developed. Oligonucleotides bearing or lacking specific binding site of the p53 protein were tail-labeled by two different modified ...
A Preliminary Study of Modification of 7-Deazaadenine with a Complex of Osmium Tetroxide with 2,2 '-Bipyridine
Vítová, Lada; Havran, Luděk; Fojta, Miroslav; Šedo, O.; Zdráhal, Z.; Vespalec, Radim
2014 - English
Reactivity of a purine nucleic base analogue 7-deazaadenine (A*) in short oligodeoxynucleotide (ODN) towards an osmium tetroxide complex with 2,2'-bipyridine (Os, bipy) was investigated by means of electrochemistry, capillary electrophoresis and MALDI-TOF mass spectrometry. Electrochemical measurement, electrophoretic analyses and mass spectrometric analyses of reaction mixture proved that ODN with an A* residue yields an osmium adduct with properties similar to those exhibited by the well-known adduct of thymine.
Keywords:
dna; bases; Electrochemical analysis
Available at various institutes of the ASCR
A Preliminary Study of Modification of 7-Deazaadenine with a Complex of Osmium Tetroxide with 2,2 '-Bipyridine
Reactivity of a purine nucleic base analogue 7-deazaadenine (A*) in short oligodeoxynucleotide (ODN) towards an osmium tetroxide complex with 2,2'-bipyridine (Os, bipy) was investigated by means of ...
Magnetic Beads-Based Electrochemical Techniques for DNA-Protein Interaction Monitoring
Fojta, Miroslav; Pivoňková, Hana; Němcová, Kateřina; Horáková Brázdilová, Petra; Havran, Luděk; Orság, Petr; Vidláková, Pavlína; Macíčková-Cahová, Hana; Balintová, Jana; Hocek, Michal
2011 - English
Electrochemical techniques, in connection with separation of nucleoprotein complexes at magnetic beads, are suitable for the monitoring of DNA-protein interactions. For the detection of complexes captured at the beads it is possible to utilize intrinsic electrochemical activity of the protein, intrinsic structure-selective signals of the DNA, or indicator DNA substrates tail-labeled with electroactive moieties.
Keywords:
DNA-protein interaction; Electrochemistry; Magnetic beads
Available at various institutes of the ASCR
Magnetic Beads-Based Electrochemical Techniques for DNA-Protein Interaction Monitoring
Electrochemical techniques, in connection with separation of nucleoprotein complexes at magnetic beads, are suitable for the monitoring of DNA-protein interactions. For the detection of complexes ...
Using Anthraquinone Labeled Nucleotide Triphosphates for Electrochemical Analysis of DNA
Vidláková, Pavlína; Balintová, Jana; Horáková Brázdilová, Petra; Havran, Luděk; Orság, Petr; Fojta, Miroslav; Hocek, Michal
2011 - English
Base-modified anthraquinone-nucleotide triphosphates were used for DNA labeling. Anthraquinone modified nucleotides were incorporated into DNA by primer extension with KOD polymerase. In cyclic and square wave voltammetry at HMDE or PGE antraquinone gave a well developed characteristic signal suitable for differentiation of the labeled DNA from unlabeled one.
Keywords:
DNA; Anthraquinone; Electrochemical analysis
Available at various institutes of the ASCR
Using Anthraquinone Labeled Nucleotide Triphosphates for Electrochemical Analysis of DNA
Base-modified anthraquinone-nucleotide triphosphates were used for DNA labeling. Anthraquinone modified nucleotides were incorporated into DNA by primer extension with KOD polymerase. In cyclic and ...
Possibility of DNA probes preparation for want of CMG
Krejčí, Jana; Bártová, Eva
2006 - Czech
The main method of cytogenetics is the DNA fluorescence in situ hybridization (DNA-FISH), where DNA probe links to the specific locus of genome. In our laboratory, DNA probes are prepared from DNA amplified by bacterial vector (PAC/BAC). The following step is exploiting Nick-translation reaction for incorporation of reporter molecule targeted for antibody conjugate with fluorescence molecule. Specificity of prepared DNA probe is increased using Cot-DNA, which suppress the repetitive sequences. Důležitou metodou cytogenetiky je DNA fluorescenční in situ hybridizace (DNA-FISH), kdy dochází ke specifické vazbě DNA sondy na určité místo genomu. V naší laboratoři jsou DNA sondy připravovány z DNA naamplifikované pomocí bakteriálního vektoru (PAC/BAC). Nicktranslační reakcí je do DNA vkládaná reportérová molekula vizualizovatelná protilátkou konjugovanou s fluorescenční látkou. Vystíněním repetitivních sekvencí pomocí Cot-DNA je následdně zvýšena specifičnost připravené sondy.
Keywords:
cytogenetics; FISH; DNA probe
Available at various institutes of the ASCR
Possibility of DNA probes preparation for want of CMG
The main method of cytogenetics is the DNA fluorescence in situ hybridization (DNA-FISH), where DNA probe links to the specific locus of genome. In our laboratory, DNA probes are prepared from DNA ...
Origin of life on Earth. Facts and (pseudo)interpretations
Paleček, Emil
2005 - Czech
In the first part there is given a summary of present knowledge on the origin of life on Earth. Further there are given several views of acknowledged scientists on science. These opinions are confronted with some interpretations of facts in popular articles. The paper warns against the danger of influencing science by ideology. V první části je uveden souhrn soudobých poznatků o vzniku života na Zemi. Dále je uvedeno několik názorů uznávaných vědců na vědu. Tyto názory jsou konfrontovány s některými interpretacemi fakt ve vědecko-populárních článcích. Přednáška upozorňuje na nebezpečí ideologizace vědy.
Keywords:
origin of life; science - interpretation
Available at various institutes of the ASCR
Origin of life on Earth. Facts and (pseudo)interpretations
In the first part there is given a summary of present knowledge on the origin of life on Earth. Further there are given several views of acknowledged scientists on science. These opinions are ...
NRGL provides central access to information on grey literature produced in the Czech Republic in the fields of science, research and education. You can find more information about grey literature and NRGL at service web
Send your suggestions and comments to nusl@techlib.cz
Provider
Other bases