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Application for the fluorescent detection of single nucleotide difference using cytosine bulge hairpin primers

Research Project

Project/Area Number 20611010
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Chemical biology
Research InstitutionOsaka University

Principal Investigator

TAKEI Fumie  Osaka University, 産業科学研究所, 助教 (30252711)

Co-Investigator(Kenkyū-buntansha) NAKATANI Kazuhiko  大阪大学, 産業科学研究所, 教授 (70237303)
Project Period (FY) 2008 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2010: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2009: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2008: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
KeywordsPCR / ヘアピンプライマー / 蛍光 / SNPタイピング / DNA / 蛍光分子 / シトシンバルジ / ヘアピン構造 / シトシンバルジ構造 / 一塩基多型
Research Abstract

PCR primer having a cytosine-bulge site at the end of 5' was designed, and using fluorescence molecule, we investigated a novel method of SNP typing. This fluorescence molecule did not inhibit on the PCR, and the hairpin unit was extended in the PCR. The fluorescence molecule binding specifically to C-bulge unit could bind to the PCR-hairpin- primer, however, after PCR, it could bind to the hairpin primer because of disappearance of C-bulge unit. The measurement of fluorescence intensity showed the PCR products, which showed this novel method could be used for the SNP typing.

Report

(4 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • 2008 Annual Research Report
  • Research Products

    (23 results)

All 2010 2009 2008 Other

All Journal Article (8 results) (of which Peer Reviewed: 8 results) Presentation (14 results) Patent(Industrial Property Rights) (1 results)

  • [Journal Article] Transformation of cytosine to uracil in single-stranded DNA via their oxime sulfonates.2010

    • Author(s)
      Oka, Y., Takei, F., Nakatani, K.
    • Journal Title

      Chem.Commun. 46

      Pages: 3378-3380

    • NAID

      130004597226

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Transformation of cytosine to uracil in single-stranded DNA via theiroxime sulfonates.2010

    • Author(s)
      Oka, Y.; Takei, F.; Nakatani, K.
    • Journal Title

      Chem.Commun.

      Volume: 46 Pages: 3378-3380

    • NAID

      130004597226

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Secondary Structure-Inducible Ligand Fluorescence Coupled with PCR2009

    • Author(s)
      Takei, F., Igarashi, M., Hagihara, M., Oka, Y., Soya, Y., Nakatani, K.
    • Journal Title

      Angew.Chem.Int.Ed. 48

      Pages: 7822-7824

    • Related Report
      2010 Final Research Report 2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Multifunctional Initiator with Platinum-Acetylide Dendritic Core for Living Polymerization of Isocyanides2009

    • Author(s)
      Onitsuka, K., Shimizu, A., Takei, F., Takahashi, S.
    • Journal Title

      Journal of Inorganic and Organometallic Polymers and Materials 19

      Pages: 98-103

    • Related Report
      2010 Final Research Report 2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthesis and Reaction of DNA Oligomers Containing Modified Cytosines Related to Bisulfite Sequencing2009

    • Author(s)
      Oka, Y., Peng, T., Takei, F., Nakatani, K.
    • Journal Title

      Org.Lett. 11

      Pages: 1377-1379

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Organoruthenium Dendrimers Possessing Tris (4-ethynylphenyl) amine Bridges2008

    • Author(s)
      Onitsuka, K., Ohara, N., Takei, F., Takahashi, S.
    • Journal Title

      Organometallics 27

      Pages: 25-27

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Transformation of cytosine to uracil in single-stranded DNA via their oxime sulfonates

    • Author(s)
      Oka, Y., Takei, F., Nakatani, K.
    • Journal Title

      Chem.Commun. (In press)

    • NAID

      130004597226

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Synthesis and Reaction of DNA Oligomers Containing Modified Cytosines Related to Bisulfite Sequencing

    • Author(s)
      Yoshimi Oka, Tao Peng, Fumie Takei, Kazuhiko Nakatani
    • Journal Title

      ORGANIC LETTERS (In press)

    • Related Report
      2008 Annual Research Report
    • Peer Reviewed
  • [Presentation] Application for the fluorescent detection of single nucleotide difference using cytosine bulge hairpin primers and competitor primers2010

    • Author(s)
      Takei F., Nakarani, K.
    • Organizer
      PACIFICHEM 2010
    • Place of Presentation
      USA
    • Year and Date
      2010-12-19
    • Related Report
      2010 Final Research Report
  • [Presentation] Application for the fluorescent detection of single nucleotide difference using cytosine bulge hairpin primers and competitor primers2010

    • Author(s)
      Fumie Takei, Masaki Hagihara, Kazuhiko Nakatani
    • Organizer
      PACIFICHEM 2010
    • Place of Presentation
      ハワイコンベンションセンター(米国)
    • Year and Date
      2010-12-17
    • Related Report
      2010 Annual Research Report
  • [Presentation] Novel Fluorescent Detection of Virus Using Cytosine Bulge Hairpin Primers for PCR2010

    • Author(s)
      Takei F.
    • Organizer
      K.Virus and Infections-2010
    • Place of Presentation
      Korea
    • Year and Date
      2010-07-31
    • Related Report
      2010 Final Research Report
  • [Presentation] Novel Fluorescent Detection of Virus Using Cytosine Bulge Hairpin Primers for PCR2010

    • Author(s)
      Fumie Takei
    • Organizer
      WCVI-2010
    • Place of Presentation
      BEXCOコンベンションセンター(韓国)
    • Year and Date
      2010-07-30
    • Related Report
      2010 Annual Research Report
  • [Presentation] ヘアピンプライマーを用いた革新的一塩基多型の蛍光検出法2010

    • Author(s)
      武井史恵、萩原正規、中谷和彦
    • Organizer
      日本ケミカルバイオロジー研究会 第5回年会
    • Place of Presentation
      慶応義塾大学(横浜市)
    • Year and Date
      2010-05-19
    • Related Report
      2010 Annual Research Report
  • [Presentation] ヘアピンタグを持つプライマー使った一塩基多型の蛍光検出2010

    • Author(s)
      武井史恵・松永静香・前田あゆみ・萩原正規・中谷和彦
    • Organizer
      日本化学会第90春季年会
    • Place of Presentation
      近畿大学 (大阪)
    • Year and Date
      2010-03-28
    • Related Report
      2009 Annual Research Report
  • [Presentation] DNA-Labeling by Secondary Structure Inducible Ligand Fluorescence2009

    • Author(s)
      Takei, F.Hagihara, M., Nakarani, K.
    • Organizer
      IKCOC-11
    • Place of Presentation
      Japan
    • Year and Date
      2009-11-09
    • Related Report
      2010 Final Research Report
  • [Presentation] ヘアピンプライマーPCR法による迅速遺伝子診断2009

    • Author(s)
      武井史恵, 中谷和彦
    • Organizer
      Crest 第5回公開シンポジウム
    • Place of Presentation
      みらいCANホール (東京都)
    • Year and Date
      2009-08-03
    • Related Report
      2009 Annual Research Report
  • [Presentation] 新規5-メチルシトシンシークエンシング2009

    • Author(s)
      岡芳美, 武井史恵, 中谷和彦
    • Organizer
      Crest 第5回公開シンポジウム
    • Place of Presentation
      みらいCANホール (東京都)
    • Year and Date
      2009-08-03
    • Related Report
      2009 Annual Research Report
  • [Presentation] DNA-Labeling by Secondary Structure Inducible Ligand Fluorescence2009

    • Author(s)
      Fumie Takei, Masako Igarashi, Masaki Hagihara, Yoshimi Oka, Yoshihiro Soya, Kazuhiko Nakatani
    • Organizer
      IKCOC-11
    • Place of Presentation
      リーガロイヤルホテル (京都)
    • Related Report
      2009 Annual Research Report
  • [Presentation] The Fluorescence Detection of Single Nucleotide Difference Using Cytosine Bulge Hairpin Primers2008

    • Author(s)
      Takei F., Hagihara, M., Nakarani, K.
    • Organizer
      XXII IUPAC SYMPOSIUM ON PHOTOCHEMISTRY 2008
    • Place of Presentation
      Sweden
    • Year and Date
      2008-07-28
    • Related Report
      2010 Final Research Report
  • [Presentation] THE FLUORESCENT DETECTION OF SINGLE NUCLEOTIDE DIFFERENCEUSING CYTOSINE BULGE HAIRPIN PRIMERS2008

    • Author(s)
      Fumie Takei, Masaki Hagihara, Yoshimi Oka, Kazuhiko Nakatani
    • Organizer
      XXII IUPAC SYMPOSIUM ON PHOTOCHEMISTRY2008
    • Place of Presentation
      SWEDEN, GOTHENBURG
    • Related Report
      2008 Annual Research Report
  • [Presentation] シトシンハルジーヘアピンプライマーを使った革新的遺伝子変異の蛍光検出2008

    • Author(s)
      武井 史恵, 岡 芳美, 萩原 正規, 曽家 義博, 中谷 和彦
    • Organizer
      日本ケミカルバイオロジー研究会 第3回年会
    • Place of Presentation
      東京
    • Related Report
      2008 Annual Research Report
  • [Presentation] The reaction of cytosine with bisulfite by base flipping from the duplex2008

    • Author(s)
      Yoshimi Oka, Tao Peng, Fumie Takei, Kazuhiko Nakatani
    • Organizer
      Joint Symposium of 18th International Roundtable on Nucleosides, Nucleotides and Nucleic Acids (IRTXVIII)
    • Place of Presentation
      Kyoto
    • Related Report
      2008 Annual Research Report
  • [Patent(Industrial Property Rights)] 一塩基多型を検出する方法および一塩基多型を検出するための試薬キット2010

    • Inventor(s)
      中谷和彦、武井史恵、萩原正規
    • Industrial Property Rights Holder
      大阪大学
    • Industrial Property Number
      2010-054658
    • Filing Date
      2010-03-11
    • Related Report
      2010 Annual Research Report 2010 Final Research Report

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Published: 2008-04-01   Modified: 2016-04-21  

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