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Development of a method of sequence library preparation for the amplification-free single-cell sequencing

Research Project

Project/Area Number 18K06195
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Review Section Basic Section 43060:System genome science-related
Research InstitutionInstitute of Physical and Chemical Research

Principal Investigator

Oguchi Yusuke  国立研究開発法人理化学研究所, 開拓研究本部, 研究員 (40599370)

Project Period (FY) 2018-04-01 – 2021-03-31
Project Status Completed (Fiscal Year 2020)
Budget Amount *help
¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
Fiscal Year 2020: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2019: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2018: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Keywordsシーケンス技術開発 / 1細胞トランスクリプトーム解析 / 1細胞トランスクリプトーム解析 / トランスクリプトーム / 機能性RNA / 遺伝子発現調節 / ゲノム解析技術 / 遺伝子ネットワーク
Outline of Final Research Achievements

In this study, we have developed a novel method of capturing a sequence library (sample) for our amplification-free sequencing system. In the conventional method, a capture probe for capturing samples is fixed to the sequence flow cell via a covalent bond. Whereas to this, we succeeded in obtaining the same level of sequencing performance as that of the covalent method, even though the capture probe was fixed via the non-covalent biotin-avidin interaction. This enables the sample to be captured by the capture probe in advance outside the flow cell, which could be advantageous for the preparation of minute amount samples such as extracted from a single cell.

Academic Significance and Societal Importance of the Research Achievements

この直近の10年において次世代シーケンサーというDNA配列を解読する装置の性能向上に伴い,生命科学・医学分野において遺伝子の発現解析が盛んに行われるようになった.市販される次世代シーケンサーは一般に,解析するサンプルをPCRによって増幅する必要がある.しかし,PCRによる増幅は,配列依存的に増幅の程度が異なり,すべてのサンプルを均一に増幅することは難しい.このような状況に対して,本研究における非増幅シーケンサーはそもそもPCRによる増幅を必要としないため,市販の大多数の装置とは一線を画す解析が可能となる.本研究成果によって,この非増幅シーケンサーの活用の幅を広げることに繋がった.

Report

(4 results)
  • 2020 Annual Research Report   Final Research Report ( PDF )
  • 2019 Research-status Report
  • 2018 Research-status Report

Research Products

(3 results)

All 2020 2019 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results,  Open Access: 1 results) Book (1 results) Remarks (1 results)

  • [Journal Article] Development of a sequencing system for spatial decoding of DNA barcode molecules at single-molecule resolution2020

    • Author(s)
      Oguchi Yusuke、Shintaku Hirofumi、Uemura Sotaro
    • Journal Title

      Communications Biology

      Volume: 3

    • DOI

      10.1038/s42003-020-01499-8

    • Related Report
      2020 Annual Research Report
    • Peer Reviewed / Open Access
  • [Book] シングルセルゲノミクス2019

    • Author(s)
      渡辺 亮、鈴木 穣
    • Total Pages
      219
    • Publisher
      羊土社
    • ISBN
      9784758103831
    • Related Report
      2019 Research-status Report
  • [Remarks] DNAバーコードの1分子空間解析 -より高い分解能での遺伝子空間分布解析が可能に-

    • URL

      https://www.riken.jp/press/2020/20201218_2/index.html

    • Related Report
      2020 Annual Research Report

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Published: 2018-04-23   Modified: 2022-01-27  

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