| Project/Area Number |
04670362
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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 | KEIO UNIVERSITY |
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Principal Investigator |
SHINOZUKA Tatsuo Keio University, Asistant Professor Legal Medicine, School of Medicine, 医学部, 講師 (70095610)
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| Co-Investigator(Kenkyū-buntansha) |
HARA Masaaki Legal Medicine, Research Assistant The Saitama Medical School, 助手 (50129160)
YANAGIDA Junichi Keio University, Professor Legal Medicine, School of Medicine, 医学部, 教授 (70049790)
HIROSE Tadaaki Keio University, Asistant Professor School of Medicine, 医学部, 講師 (60051405)
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| Project Period (FY) |
1992 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | DNA polymorphism / Paternity / Restriction Fragment Length Polymorphisms (RFLPs) / Polymerase Chain Reaction (RCR) / HLA DQ alpha Typing / RFLPs法 / HLADQalpha型 / RFLP法 / HLADQα |
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| Research Abstract |
DNA was extracted from cryopreserved lymphocytes after paternity testing by the phenol-chloroform method or commercial DNA extraction kits. Paternity analysis has been performed by a simple method for nonradioactive DNA fingerprinting. The method utilizes a minisatelite probe MZ1.3 labeled with digoxigenin, which are detected using an anti-digoxigenin-alkaline phosphatase complex. This system detected DNA restriction fragments in more than 10mug of DNA from lymphocytes.
On the other hand, the experiment has been performed to evaluate typing of the HLA DQ a gene by polymerase chain reaction (PCR) amplification of the gene and subsequent hybridization with sequence-specific oligonucleotide probe (Ampli Type kit). The sensitivity to detect DNA extraction from the lymphocytes was evaluated by amplifying various amounts of purified DNA.HLA DQ a typing were detected from 1-20ng as amounts of template DNA from paternity test.
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