2020 Fiscal Year Annual Research Report
Efficient polymerase chain reaction from single DNA molecules
| Project/Area Number |
18K14260
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| Research Institution | Japan Agency for Marine-Earth Science and Technology |
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Principal Investigator |
張 翼 国立研究開発法人海洋研究開発機構, 超先鋭研究開発部門(超先鋭研究プログラム), 研究員 (40795358)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Keywords | Restriction enzyme / Electrophoresis / Digital counting / Droplet / Biosensor |
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| Outline of Annual Research Achievements |
Sensitivity improvement is one of the most central tasks in the development of bioanalytical methods. After publishing the unique droplet technology last year (Zhang et al., Science Advances, 2019, 5(8): eaav8185), an unexpected experimental result allowed an unplanned R&D of a new method to quantify the digest efficiency of any kinds of restriction enzymes, a new application of digital bioassays, with unprecedented sensitivity. Once I used a DNA digest solution as the source of template DNA for the cell-free fluorescent protein synthesis; but surprisingly, many fluorescent droplets appeared on the array. The complete digestion was pre-confirmed using gel electrophoresis, a gold-standard confirmation experiment in every molecular biology laboratory, which suggested that the droplet system can be diverted into a convenient analytical device to interrogate the digest efficiency. And importantly, this approach quantified the template DNA of much lower concentrations that cannot be detected by the ensemble-based methods. The same rationale was extended to multiplexed assays and applicable to any DNA-degrading or genome-editing enzymes. The accumulation of these results produced a new paper this year (Zhang et al., PLOS ONE, 2020, 15(12): e0244464).
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| Remarks |
Blue Earth海と地球の情報誌, 第32巻 第3号 (通巻165号), pp. 1-3.
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