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A Rapid Digital PCR System with a Pressurized Thermal Cycler

We designed a silicon-based fast-generated static droplets array (SDA) chip and developed a rapid digital polymerase chain reaction (dPCR) detection platform that is easy to load samples for fluorescence monitoring. By using the direct scraping method for sample loading, a droplet array of 2704 micr...

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Detalles Bibliográficos
Autores principales: Chen, Xuee, Song, Qi, Zhang, Beini, Gao, Yibo, Lou, Kai, Liu, Yiteng, Wen, Weijia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8708658/
https://www.ncbi.nlm.nih.gov/pubmed/34945412
http://dx.doi.org/10.3390/mi12121562
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author Chen, Xuee
Song, Qi
Zhang, Beini
Gao, Yibo
Lou, Kai
Liu, Yiteng
Wen, Weijia
author_facet Chen, Xuee
Song, Qi
Zhang, Beini
Gao, Yibo
Lou, Kai
Liu, Yiteng
Wen, Weijia
author_sort Chen, Xuee
collection PubMed
description We designed a silicon-based fast-generated static droplets array (SDA) chip and developed a rapid digital polymerase chain reaction (dPCR) detection platform that is easy to load samples for fluorescence monitoring. By using the direct scraping method for sample loading, a droplet array of 2704 microwells with each volume of about 0.785 nL can be easily realized. It was determined that the sample loading time was less than 10 s with very simple and efficient characteristics. In this platform, a pressurized thermal cycling device was first used to solve the evaporation problem usually encountered for dPCR experiments, which is critical to ensuring the successful amplification of templates at the nanoliter scale. We used a gradient dilution of the hepatitis B virus (HBV) plasmid as the target DNA for a dPCR reaction to test the feasibility of the dPCR chip. Our experimental results demonstrated that the dPCR chip could be used to quantitatively detect DNA molecules. Furthermore, the platform can measure the fluorescence intensity in real-time. To test the accuracy of the digital PCR system, we chose three-channel silicon-based chips to operate real-time fluorescent PCR experiments on this platform.
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spelling pubmed-87086582021-12-25 A Rapid Digital PCR System with a Pressurized Thermal Cycler Chen, Xuee Song, Qi Zhang, Beini Gao, Yibo Lou, Kai Liu, Yiteng Wen, Weijia Micromachines (Basel) Article We designed a silicon-based fast-generated static droplets array (SDA) chip and developed a rapid digital polymerase chain reaction (dPCR) detection platform that is easy to load samples for fluorescence monitoring. By using the direct scraping method for sample loading, a droplet array of 2704 microwells with each volume of about 0.785 nL can be easily realized. It was determined that the sample loading time was less than 10 s with very simple and efficient characteristics. In this platform, a pressurized thermal cycling device was first used to solve the evaporation problem usually encountered for dPCR experiments, which is critical to ensuring the successful amplification of templates at the nanoliter scale. We used a gradient dilution of the hepatitis B virus (HBV) plasmid as the target DNA for a dPCR reaction to test the feasibility of the dPCR chip. Our experimental results demonstrated that the dPCR chip could be used to quantitatively detect DNA molecules. Furthermore, the platform can measure the fluorescence intensity in real-time. To test the accuracy of the digital PCR system, we chose three-channel silicon-based chips to operate real-time fluorescent PCR experiments on this platform. MDPI 2021-12-15 /pmc/articles/PMC8708658/ /pubmed/34945412 http://dx.doi.org/10.3390/mi12121562 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chen, Xuee
Song, Qi
Zhang, Beini
Gao, Yibo
Lou, Kai
Liu, Yiteng
Wen, Weijia
A Rapid Digital PCR System with a Pressurized Thermal Cycler
title A Rapid Digital PCR System with a Pressurized Thermal Cycler
title_full A Rapid Digital PCR System with a Pressurized Thermal Cycler
title_fullStr A Rapid Digital PCR System with a Pressurized Thermal Cycler
title_full_unstemmed A Rapid Digital PCR System with a Pressurized Thermal Cycler
title_short A Rapid Digital PCR System with a Pressurized Thermal Cycler
title_sort rapid digital pcr system with a pressurized thermal cycler
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8708658/
https://www.ncbi.nlm.nih.gov/pubmed/34945412
http://dx.doi.org/10.3390/mi12121562
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