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Publication detail
Gaňová Martina, Neužil Pavel
Original Title
Development of digital polymerase chain reaction
Type
abstract
Language
English
Original Abstract
During the last 20 years, microfluidics has proven to be a powerful nucleic acid (NA) analysis tool in applications such as point-of-care diagnostic methods, which critically depended on miniaturization. From simple systems they evolved into complex devices integrating sample processing, NA amplification and product detection using samples with an ultra-small volume, resulting in inexpensive screening. There are even techniques which cannot be conducted without microfluidics, such as digital polymerase chain reaction (dPCR) developed in 1999. We demonstrate a concept of droplet real-time PCR (qPCR) which will be developed into dPCR. For droplet real-time detection, the amplification is conducted in 0.3-µL master mixture droplet containing target gene encapsulated in 2 µL of mineral oil. For digital detection, a chip-based system with microwell sample dispersion will be developed. The 3 µL of master mix solution with DNA will be pipetted on a silicone chip and covered with a hydrophobically treated glass. We improved the features of both devices where the experimental setup for qPCR and dPCR is the same, except the detectors. Our detection system is more economic thanks to the use of a recyclable microchip, lower consumption of components and shortened detection time, which makes it attractive for point-of-care applications.
Keywords
Digital PCR, miniaturization, microchip technology
Authors
Released
26. 2. 2021
Publisher
TAYLOR & FRANCIS LTD
Location
European Biotechnology Congress 2020 Abstracts, ABINGDON
ISBN
1314-3530
Periodical
Biotechnology and Biotechnological Equipment
Year of study
35
Number
sup1
State
United Kingdom of Great Britain and Northern Ireland
Pages from
S62
Pages to
S128
Pages count
67
URL
https://doi.org/10.1080/13102818.2020.1871545