| Project/Area Number |
05807174
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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 |
Functional basic dentistry
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| Research Institution | TOKYO DENTAL COLLEGE |
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Principal Investigator |
HANAOKA Yoichi Tokyo Dental College, Forensic Odontology, Assistant professor, 歯学部法歯学講座, 講師 (30180912)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1994: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1993: ¥800,000 (Direct Cost: ¥800,000)
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| Keywords | Tooth / DNA analysis / DNA finger priniting / D4S43 locus / Sex determination / ASF領域 / 歯牙 / PCR増幅 / DNA抽出 / 歯髄 / 歯牙の脱灰 |
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| Research Abstract |
1. Extraction of DNA from tooth : The amount of DNA obtained from the dental pulp tissue of single tooth varied from 3-40mug and high molecular weight of DNA was still present in samples stored about for a year. No correlation was observed between the storage period of the tooth samples and the DNA extraction ratio, but when the dental pulp tissue was more than 5mg in weight the amount of DNA extracted was more than 10mug. In samples of tooth hard tissues decalcification for a week using 0.5M EDTA solution was useful for extraction of DNA and in most cases, recovered DNA from tooth hard tissues was degraded.
2. Analysis of polymorphism using extracted DNA from tooth : when the high molecular weight of DNA was recovered DNA finger printing is useful and finger print patterns obtained from DNA extracted from tooth and blood samples were identical. However DNA finger printing needs 3-5mug high molecular weight of DNA and was not suitable for DNA recovered from tooth hard tissues. The DNA recovered from tooth hard tissues was suitable only for PCR analysis.
3. VNTR analysis in D4S43 locus by PCR : VNTR polymorphism at D4S43 of Japanese polulation was detected using blood samples before using tooth samples. The largest allele frequency was 60% (184bp range) and the hetero zygosity value calculated to be 59%. In addition, four types' size variation was found in the band on 184bp that was reported including one 14bp core in repeated region of D4S43 locus as other kind of polymorphism.
4. Sex determination using DNA obtained from tooth : Sex determination by Witt & Erickson's method was succeeded with dual PCR and restriction enzyme digestion. However about 14% of male samples of tooth hard tissues were not able to determined.
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