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A Sensitive Method of DNA Fingerprinting Using Biotinylated RNA Probe

Research Project

Project/Area Number 01570340
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Legal medicine
Research InstitutionUniversity of the Ryukyus

Principal Investigator

NAGAMORI Hajime  University of the Ryukyus, School of Medicine, Department of Legal Medicine, Professor, 医学部, 教授 (50045605)

Co-Investigator(Kenkyū-buntansha) NAGAMINE Masaru  University of the Ryukyus, School of Medicine, Second Department of Biochemistry, 医学部, 助手 (20189161)
KAJIWARA Masahiro  Kyorin University, School of Medicine, Department of Legal Medicine, Instructor, 医学部, 助手 (30152656)
UCHIMA Eikoh  University of the Ryukyus, School of Medicine, Department of Legal Medicine, Ins, 医学部, 助手 (90093377)
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1990: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsIndividual identification / Paternity testing / DNA polymorphism / Southern hybridization / DNA fingerprinting / Biotinylated probe / DNA指紋法 / サザントランスファ-
Research Abstract

Jeffreys and his colleagues have developed a DNA fingerprinting system utilizing a variable number of tandem repeats. They produced minisatellite probes which were able to simultaneously hybridize to a number of highly variable DNA regions. The Southern hybridization patterns obtained with these probes consist of multiple hypervariable fragments and are specific to individuals.
DNA fingerprinting has become a powerful new method for forensic applications including individual identification and paternity testing. This method is restrictive in its application to routine work because an isotopically labeled probe is required. Thus we attempted the application of a non-radioactive biotinylated RNA probe derived from SP6 system. However, we have failed to obtain clear fingerprinting due to the lack of sufficient sensitivity. So we have changed another labeling method. The 33mer oligonucleotide corresponding to the repeat unit of the human myoglobin minisatellite was synthesized by the solid phase phosphoramidite method. Biotin was covalently attached via hexylamine linker like an additional base to the 5'end. DNA fragments digested with restriction enzyme were electrophoresed, blotted on nitrocellulose membrane, hybridized with the biotinylated oligonucleotide probe and washed under mild condition. A commercially available kit was used for DNA detection. It was proved that this fingerprinting was comparable to the detection by radioactive method. Therefore, this method may be available for paternity testing and forensic application.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (3 results)

All Other

All Publications (3 results)

  • [Publications] 永盛 肇,内間 栄行,長領 勝,梶原 正弘,森永 泰正,向井 敏二,大城 尚伸,大野 曜吉: "ビオチン化標識オリゴヌクレオチドプロ-ブを用いたDNAフィンガ-プリント" 法医学の実際と研究. 33. 17-20 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Hajime Nagamori, Eikoh Uchima, Masaru Nagamine, Masahiro Kajiwara, Yasumasa Morinaga, Toshiji Mukai, Takanobu Ohshiro and Youkichi Ohno: "DNA Fingerprinting Using Biotinylated Oligonucleotide Probe" The Research and Practice in Forensic Medicine. 33. 17-20 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 永盛 肇・内間 栄行・長嶺 勝・梶原 正弘・森永 泰正・向井 敏二・大城 尚伸・大野 曜吉: "ビオチン化標識オリゴヌクレオチドプロ-ブを用いたDNAフィンガ-プリント" 法医学の実際と研究. 33. 17-20 (1990)

    • Related Report
      1990 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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