On-chip Biotechnologiesが、「International Symposium on Plasmid Biology 2024」にブース出展致します。9/3 (火) 12:30にはランチョンセミナーに静岡大学の新谷 政己教授をお招きし、ご講演いただきます。
講演タイトルは「Single-cell analysis of ‘plasmid holders’ in microbial communities」。
ご興味のある方は是非ご参加くださいませ！
【開催時間】12:30-13:30
【開催場所】Room #41
【セミナー要旨】
Single-cell analysis of ‘plasmid holders’ in microbial communities
Masaki Shintani*1,2
*1 Shizuoka University, Japan; *2 Japan Collection of Microorganisms, RIKEN BioResource Research Center, Japan.
Understanding which bacteria in the environment contain specific plasmids is crucial for comprehending microbial ecology and gene transfer dynamics. Various methods have been used to address this issue, including Fluorescence in situ hybridization (FISH) or in situ PCR at the single-cell level. In our study, we developed a culture-independent, single-cell analysis technique by combining droplet digital PCR (ddPCR) with 30 µm diameter water-in-oil droplets and targeted single-cell dispensing methods using the On-chip® Droplet Selector from On-chip Biotechnologies.
We applied this innovative approach to microorganisms extracted from lake sediments, encapsulating each cell within individual droplets. Multiplex ddPCR was then performed using specific primers and a green-fluorescent probe to amplify and detect the replication initiation protein gene (repA) of the plasmid, along with primers for amplifying partial 16S rRNA gene (V3-V4). Analysis of more than 2 million droplets revealed over 500 exhibited green fluorescence, indicating the presence of cells harboring the targeted plasmid. Subsequent sorting and sequencing of the droplets led to the identification of partial 16S rRNA gene sequences, revealing several bacterial genera as potential original hosts of the plasmid.
These findings demonstrate the power of this advanced single-cell technique in determining which microbes have which plasmids, providing valuable insight into the dynamics of plasmids and their ecological role within microbial communities.
展示ブースでは、弊社製品Droplet Selectorを展示予定。
ご来場お待ち申し上げております。