Development of single-cell whole genome amplification method for uncultivable bacteria using droplet microfluidics

2015 Fiscal Year Final Research Report

• Project/Area Number: 26820365
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Waseda University
• Principal Investigator: Masahito Hosokawa 早稲田大学, ナノ・ライフ創新研究機構, 助教 (60722981)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: シングルセル / 全ゲノム増幅 / マイクロ流体デバイス / １細胞解析

Outline of Final Research Achievements

In this study, we aimed to develop a microfluidic droplet-based whole genome amplification method for single bacterial cells. We developed a microfluidic device for generation of picoliter-sized droplet for compartmentalized reaction environment. Single bacterial cells or their genome DNA fragments were encapsulated into each droplet. By compartmenting genome fragments into individual droplets, DNA fragments are amplified uniformly, that minimizes amplification bias and the effect of contaminating DNA. Our results demonstrated the potential of microfluidic generated droplets as an efficient tool for the analysis of single cell genome along with great reduction in the cost and labor investment required for the investigation of genome diversity of uncultivable bacteria.

Free Research Field

プロセス工学、生物機能・バイオプロセス