The determination of DNA melting temperature by microchip digital polymerase chain reaction

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216305%3A26620%2F21%3APU142851" target="_blank" >RIV/00216305:26620/21:PU142851 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

The determination of DNA melting temperature by microchip digital polymerase chain reaction

Popis výsledku v původním jazyce

The double-stranded (ds) structure gives deoxyribonucleic acid (DNA) physical and chemical properties that make the molecule of DNA very stable. Although, non-physiological temperature disrupts the dsDNA macromolecules and cause denaturation. The double helix DNA molecules dissociate into two single-strands at specific melting temperature (Tm) at which 50% molecules are denatured. We demonstrated the use of dsDNA as a temperature sensor for temperature calibration of our experimental digital polymerase chain reaction cycler (dPCR). DNA Tm was evaluated by monitoring fluorescence signal during heating known as melting curve analysis (MCA). The synthetic virus DNA was amplified with a commercial cycler and specific Tm was determined at 87 °C. The temperature-dependent dissociation of dsDNA was measured using a DNA intercalating fluorescence Evagreen dye. Calibration of experimental dPCR cycler was performed also by fluorescence signal detection. A LabVIEW-based temperature control program with thermoele

Název v anglickém jazyce

The determination of DNA melting temperature by microchip digital polymerase chain reaction

Popis výsledku anglicky

The double-stranded (ds) structure gives deoxyribonucleic acid (DNA) physical and chemical properties that make the molecule of DNA very stable. Although, non-physiological temperature disrupts the dsDNA macromolecules and cause denaturation. The double helix DNA molecules dissociate into two single-strands at specific melting temperature (Tm) at which 50% molecules are denatured. We demonstrated the use of dsDNA as a temperature sensor for temperature calibration of our experimental digital polymerase chain reaction cycler (dPCR). DNA Tm was evaluated by monitoring fluorescence signal during heating known as melting curve analysis (MCA). The synthetic virus DNA was amplified with a commercial cycler and specific Tm was determined at 87 °C. The temperature-dependent dissociation of dsDNA was measured using a DNA intercalating fluorescence Evagreen dye. Calibration of experimental dPCR cycler was performed also by fluorescence signal detection. A LabVIEW-based temperature control program with thermoele

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

21002 - Nano-processes (applications on nano-scale); (biomaterials to be 2.9)

Projekt

<a href="/cs/project/LM2018110" target="_blank" >LM2018110: Výzkumná infrastruktura CzechNanoLab</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2021

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů