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

Result code in 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>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

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

Original language description

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

Czech name

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Czech description

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Type

O - Miscellaneous

CEP classification

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OECD FORD branch

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

Project

<a href="/en/project/LM2018110" target="_blank" >LM2018110: CzechNanoLab research infrastructure</a><br>

Continuities

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

Publication year

2021

Confidentiality

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