DNA Fingerprinting by Means of a Nonradioactive Probe for Individual Identification
| Project/Area Number |
63570270
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Legal medicine
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| Research Institution | Niigata University |
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Principal Investigator |
YAMANOUCHI Haruo Niigata University, School of Medicine, Professor, 医学部, 教授 (30134919)
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| Co-Investigator(Kenkyū-buntansha) |
NAITO Emiko Niigata University, School of Medicine, Assistant, 医学部, 助手 (80018811)
KOMINAMI Ryo Niigata University, School of Medicine, Professor, 医学部, 教授 (40133615)
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| Project Period (FY) |
1988 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1990: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1989: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1988: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | DNA fingerprint / Nonradioactive probe / Enhanced chemiluminesent / Paternity testing / Zygosity diagnosis / Bone marrow engraftment / myoプロ-ブ / DNAフィンガ-プリンティング / コ-ルド法 / DNAフィンガープリント / Nyoプローブ / 非放射性DNAプローブ / スルフォン化標識プローブ |
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| Research Abstract |
A nonradioactive DNA fingerprinting using ECL (Enhanced Chemi-Luminesent) technique was performed. Three to five mug of genomic DNA digested with Hae III was electrophoresed in 1.3% agarose and transferred onto nylon membrane. Following alkali fixation, the blot was hybridized with peroxidase-labeledminisatellite " myo " probe and washed. Blue light araising from ECL reaction catalyzed by the enzyme label was detected on a light sensitive film. A commercially available kit was employed for DNA detection and labeling. Another minisatellite probe could be reprobed without removing myo. This technique had comparable sensitivity to radioactive method in DNA detection, and was far easier to reprobe than other non-isotopic methods using sulfonated, hapten-labeled and biotin-labeled DNA probe.
Actually, this method could be applied to paternity testing, zygosity diagnosis and confirmation of bone marrow engraftment. Thus, since the handling of cold probe is safe and not contaminable, this technique is considered feasible for routine work involving individual identification.
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Report
(4 results)
Research Products
(13 results)
