Revealing the Secrets of PCR
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216305%3A26620%2F19%3APU132890" target="_blank" >RIV/00216305:26620/19:PU132890 - isvavai.cz</a>
Result on the web
<a href="https://www.sciencedirect.com/science/article/pii/S0925400519311232" target="_blank" >https://www.sciencedirect.com/science/article/pii/S0925400519311232</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.snb.2019.126924" target="_blank" >10.1016/j.snb.2019.126924</a>
Alternative languages
Result language
angličtina
Original language name
Revealing the Secrets of PCR
Original language description
The polymerase chain reaction (PCR) method and its quantitative clone (qPCR) were a landmark discovery for detection and quantification of small amounts of unambiguous deoxyribonucleic acids (DNA) due to its enormous sensitivity and specificity. The current methods for qPCR protocol optimization provide no information of the PCR propagation during the cycles as only single data point is extracted at the end of each thermal cycles, limiting a thorough understanding of reaction details. In this study, we utilized the continuous fluorescence monitoring (CFM) method to observe progress of the reaction in real time with over 100 data points per each cycle, thus gaining a profound insight into the PCR itself. This provided information about the real-time PCR status, dominating reactions and their completion/incompletion during each cycle as well as their reaction kinetics. We then adjusted the duration of either annealing or elongation steps to ensure their completion within each cycle, resulting in the protocol optimization with complete amplification, enhancing PCR efficiency and taking < 20 min to obtain maximum product amount. The proposed method was verified using DNA with lengths of 177 bp, 250 bp, and 400 bp. It can also be adopted for helping with qPCR troubleshooting as well as protocol optimizing just by reprogramming commercial real-time PCR cycler.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
20602 - Medical laboratory technology (including laboratory samples analysis; diagnostic technologies) (Biomaterials to be 2.9 [physical characteristics of living material as related to medical implants, devices, sensors])
Result continuities
Project
<a href="/en/project/GA16-11140S" target="_blank" >GA16-11140S: Microfludics-based Ultra Fast Differential Scanning Fluorimetry for Drug Discovery (µDSF)</a><br>
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2019
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Sensors and Actuators B: Chemical
ISSN
0925-4005
e-ISSN
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Volume of the periodical
298
Issue of the periodical within the volume
NA
Country of publishing house
CH - SWITZERLAND
Number of pages
6
Pages from-to
1-6
UT code for WoS article
000483419800024
EID of the result in the Scopus database
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