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Info. Vol.16 - No.4 (2022.12.20)
Title Micro-injection Molded Droplet Generation System for Digital PCR Application
Authors Daae Jo1, So Young Kim2,3, Hyeon Woo Kang2,3, Sung Han Pyo1, Nam Kyu Jeong1, Nam ho Bae1, Seok Jae Lee1, Yong Tae Kim4, Kyoung G. Lee1

*Seok Jae Lee sjlee@nnfc.re.kr
*Yong Tae Kim ytkim@tukorea.ac.kr
*Kyoung G. Lee kglee@nnfc.re.kr
Institutions 1National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 34141, Republic of Korea
2Bio R&D Lab, BioTNS Co.Ltd, 19-12 Daehak-ro 76 beonan-gil, Daejeon 34141, Republic of Korea
3Department of Biology, Soon Chun Hyang University, 22 Soonchunhyang-ro, Chungcheongnam-do, Asan-si 31538, Republic of Korea
4Department of Chemical Engineering and Biotechnology, Tech University of Korea, 237 Sangidaehak-ro, Siheung-si, Gyeonggi-do 15073, Republic of Korea
Abstract Sensitive, effective, and quantitative analysis of infectious pathogens is an important task for the prevention of human health threats. Herein, we present an advanced approach to producing gene-encapsulated microdroplets for quantitative analysis using a micropatterned metal mold and injection molding technique with an automatically operated system. An injection molded microdroplet generation device was successfully fabricated with a minimum channel width of 30 μm and optimized to produce 100 μm diameter droplets. The optimized microchannel design and flow rate also enable the production of stable numbers of microdroplets (~ 16, 000 droplets). To verify the applicability of our device and system to droplet-based digital PCR analysis, Escherichia coli ( E. coli ) O157:H7 was selected as a model bacterial pathogen, and the stx2 gene was amplified in the microdroplets. The generated microdroplets exhibit both chemical and mechanical stability, and our results are similar to those obtained by a commercially available method. Accordingly, the usefulness of the microdroplet generative device and system is confirmed as a simple, fast, and reliable tool for the quantitative molecular analysis of infectious diseases.
Keyword Digital nucleic acid assay, Microdroplet, Micropattern, Digital PCR, Metal mold
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