Efficient polymerase chain reaction from single DNA molecules

2020 Fiscal Year Final Research Report

• Project/Area Number: 18K14260
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 34020:Analytical chemistry-related
• Research Institution: Japan Agency for Marine-Earth Science and Technology
• Principal Investigator: Zhang Yi 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)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: High throughput / Microfabrication / Single molecule / Enzyme / Sensitivity / Poisson statistics / Image processing / Screening

Outline of Final Research Achievements

We cannot study what we cannot measure. This simple philosophy is the basic driving force for analytical chemistry to move forward. This Early-Career Scientists study was also planned and implemented based on this general interest. My research platform features high throughput and deals with a massive number of single DNA molecules, which enabled: (1) cell-free protein synthesis from every single DNA molecule; (2) genotype-phenotype linkage at the single DNA molecule; (3) sorting and amplification for single DNA molecules; and (4) amplification-free single DNA molecule detection. During the period of this research, I succeeded in obtaining the world's most active enzyme capable of hydrolyzing phosphate esters, establishing the novel digital high-throughput screening theory, and developing the world's most sensitive method capable of evaluating the performance of any DNA-cleaving enzymes.

Free Research Field

Analytical chemistry

Academic Significance and Societal Importance of the Research Achievements

The massively parallel, highly uniform, and individually addressable femtoliter droplet array technology offered a high-end standard and new possibilities for future diverse bioanalytical applications. The integrated system also stressed the significance of the multidisciplinary approach.