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An integrated digital PCR system with high universality and low cost for nucleic acid detection
Wang, Kangning1; Li, Bin2; Guo, Yu3; Wu, Yanqi4; Li, Yan1; Wu, Wenming1
2022-08-19
Source PublicationFrontiers in Bioengineering and Biotechnology
Volume10
Abstract

Digital PCR is the most advanced PCR technology. However, due to the high price of the digital PCR analysis instrument, this powerful nucleic acid detection technology is still difficult to be popularized in the general biochemistry laboratory. Moreover, one of the biggest disadvantages of commercial digital PCR systems is the poor versatility of reagents: each instrument can only be used for a few customized kits. Herein, we built a low-cost digital PCR system. The system only relies on low-cost traditional flat-panel PCR equipment to provide temperature conditions for commercial dPCR chips, and the self-made fluorescence detection system is designed and optically optimized to meet a wide range of reagent requirements. More importantly, our system not only has a low cost (<8000 US dollars) but also has a much higher universality for nucleic acid detection reagents than the traditional commercial digital PCR system. In this study, several samples were tested. The genes used in the experiment were plasmids containing UPE-1a fragment, TP53 reference DNA, hepatitis B virus DNA, leukemia sample, SARS-COV-2 DNA, and SARS-COV-2 RNA. Under the condition that DNA can be amplified normally, the function of the dPCR system can be realized with simpler and low-price equipment. Some DNA cannot be detected by using the commercial dPCR system because of the special formula when it is configured as the reaction solution, but these DNA fluorescence signals can be clearly detected by our system, and the concentration can be calculated. Our system is more applicable than the commercial dPCR system to form a new dPCR system that is smaller and more widely applicable than commercially available machinery.

KeywordAbsolute Quantification Digital Pcr Dpcr Chip Fluorescence Detection Sars-cov-2 Virus
DOI10.3389/fbioe.2022.947895
URLView the original
Language英語English
WOS Research AreaBiotechnology & Applied Microbiology ; Science & Technology - Other Topics
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000849740000001
Scopus ID2-s2.0-85137149161
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Document TypeJournal article
CollectionUniversity of Macau
Affiliation1.Institute of Biological and Medical Engineering, Guangdong Academy of Sciences, Guangzhou, China
2.Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
3.School of Mechanical and Electrical Engineering, Guangdong University of Technology, Guangzhou, China
4.State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Taipa, Macao
Recommended Citation
GB/T 7714
Wang, Kangning,Li, Bin,Guo, Yu,et al. An integrated digital PCR system with high universality and low cost for nucleic acid detection[J]. Frontiers in Bioengineering and Biotechnology, 2022, 10.
APA Wang, Kangning., Li, Bin., Guo, Yu., Wu, Yanqi., Li, Yan., & Wu, Wenming (2022). An integrated digital PCR system with high universality and low cost for nucleic acid detection. Frontiers in Bioengineering and Biotechnology, 10.
MLA Wang, Kangning,et al."An integrated digital PCR system with high universality and low cost for nucleic acid detection".Frontiers in Bioengineering and Biotechnology 10(2022).
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