| 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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