| 研究実績の概要 |
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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