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

2010 Fiscal Year Final Research Report

• Project/Area Number: 20611010
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemical biology
• Research Institution: Osaka 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
• Keywords: PCR / ヘアピンプライマー / 蛍光 / SNPタイピング / DNA

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.

Research Products

• [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
  • Peer Reviewed
• [Journal Article] Secondary Structure-Inducible Ligand Fluorescence Coupled with PCR (2009)
  • Author(s): Takei, F., Igarashi, M., Hagihara, M., Oka, Y., Soya, Y., Nakatani, K.
  • Journal Title: Angew.Chem.Int.Ed. 48 Pages: 7822-7824
  • Peer Reviewed
• [Journal Article] Multifunctional Initiator with Platinum-Acetylide Dendritic Core for Living Polymerization of Isocyanides (2009)
  • Author(s): Onitsuka, K., Shimizu, A., Takei, F., Takahashi, S.
  • Journal Title: Journal of Inorganic and Organometallic Polymers and Materials 19 Pages: 98-103
  • Peer Reviewed
• [Journal Article] Synthesis and Reaction of DNA Oligomers Containing Modified Cytosines Related to Bisulfite Sequencing (2009)
  • Author(s): Oka, Y., Peng, T., Takei, F., Nakatani, K.
  • Journal Title: Org.Lett. 11 Pages: 1377-1379
  • Peer Reviewed
• [Journal Article] Organoruthenium Dendrimers Possessing Tris (4-ethynylphenyl) amine Bridges (2008)
  • Author(s): Onitsuka, K., Ohara, N., Takei, F., Takahashi, S.
  • Journal Title: Organometallics 27 Pages: 25-27
  • Peer Reviewed
• [Presentation] Application for the fluorescent detection of single nucleotide difference using cytosine bulge hairpin primers and competitor primers (2010)
  • Author(s): Takei F., Nakarani, K.
  • Organizer: PACIFICHEM 2010
  • Place of Presentation: USA
  • Year and Date: 2010-12-19
• [Presentation] Novel Fluorescent Detection of Virus Using Cytosine Bulge Hairpin Primers for PCR (2010)
  • Author(s): Takei F.
  • Organizer: K.Virus and Infections-2010
  • Place of Presentation: Korea
  • Year and Date: 2010-07-31
• [Presentation] DNA-Labeling by Secondary Structure Inducible Ligand Fluorescence (2009)
  • Author(s): Takei, F.Hagihara, M., Nakarani, K.
  • Organizer: IKCOC-11
  • Place of Presentation: Japan
  • Year and Date: 2009-11-09
• [Presentation] The Fluorescence Detection of Single Nucleotide Difference Using Cytosine Bulge Hairpin Primers (2008)
  • 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
• [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: 特許,特願2010-054658
  • Filing Date: 2010-03-11