• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.497-513
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• 1995

Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.515-528
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• 1995

The human genomic deoxyribonucleic acid(DNA) was extracted from the pulp, dentin of 22 teeth by clelex, phenol methods. Samples of the tooth-derived DNA were amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologus amelogenin gene and D1S80 locus detection The following results have been achieved. 1. Chelex and phenol method are effective to sex determination in the pulp and dentin 2. Chelex method is not suitable for detection of D1S80 locus. 3. Concentration and purity of DNA for teeth using chelex method is lower than using phenol method. From the above investigation, chelex method is simple, rapid for sex determination, but it is not suitable for detection of VNTRs.

• Journal of Oral Medicine and Pain
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• v.19 no.2
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• pp.205-219
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• 1994

Cigarette butts from 5 smokers were gathered and then, placed in room temperature for 1, 3, 5, 7, 15 days. The possible use of the cigarette butts for individual identification was evaluated in sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene from the extracted DNA. 1. DNA extraction was possible in cigarette butts weree left in room temperature for 15days, so it can be applicatable to individual identification by polymerase chain reaction(PCR). 2. Amplification of X-Y homologous amelogenin gene by PCR made it possible to identify the sex in saliva stains (cigarette butts). 3. Amplification of D1S80 locus can be acquired from adding the boving serum albumin and hot start PCR procedures from forensic samples such as saliva stains (cigarette butts), so the AMP-FLPs examining is possible. 4. Genotype could be determined simply and rapidly using Amplitype$TM$ HLA-DQ$\alpha$ forensic kit in examining the HLA-DQA1 gene. From the investigation, DNA extraction, sex determination, amplification of D1S80 locus, polymorphisms of HLA-DQA1 gene was successfully done even though the cigarette butts were left for 15 days at room temperature. Therefore cigarette butts are highly reliable and applicatable as molecular biologic samples for individual identification.

• Journal of Oral Medicine and Pain
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• v.20 no.1
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• pp.229-246
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• 1995

The deoxyribonucleic acid(DNA) was isolated from the pulp, dentin and enamel of the 4 fresh teeth and the 7 teeth left in room temperature for 10 years. Then it was examed to find out the usefulness for forensic dental medicine. Samples of the tooth-derived DNA amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologous amelogenin gene and D1S80 locus detection. The following results have been achieved. DNA extraction was possible in pulp and dentin of the fresh teeth, so it could be applicatable to detection of X-Y homologous amelogenin gene for sex determination, amplification of D1S80 locus by PCR. Sex determination was possible in pulp and dentin of the teeth left at room temperature for 10 years. Also, possible the detection fo AMP-FLPs to increase PCR cycling up to 40. DNA was isolated from all pulp of the fresh teeth and the teeth left in room temperature for 10 years, and also isolated from the dentin of the fresh teeth, partially isolated(3/7) from the dentin of the teeth left in room temperature for 10 years, but DNA was not isolated from enamel. From the above investigation, DNA extraction, sex determination, amplification of D1S80 locus were successfully accomplished even though the teeth were left for 10 years at room temperature. Therefore, teeth, especially pulp, are highly reliable and applicable as molecular biological samples for individual identification.

• BMB Reports
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• v.28 no.4
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• pp.359-362
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• 1995

The highly polymorphic variable number of tandem repeat (VNTR) loci in the human genome are informative markers for the genetic characterization of individuals in the paternity test and forensic science as well as for the study of human disease. In this study, VNTR loci D1S80 and D2S123 have been amplified by PCR and the amplified length polymorphic alleles were detected with a discontinuous vertical PAGE system and silver staining. For explicit DNA typing, PCR optimization, in which amplification efficiencies are similar over a wide range of allele sizes, non-specific amplifications are minimal, and new longer alleles have high amplification efficiency, has been performed by changing the PCR reaction buffer composition and thermal cycling conditions. It turned out that adding an appropriate amount of Tween 20 and NP40 to the PCR reaction buffer and raising the annealing temperature to $68^{\circ}C$ in thermal cycling made it possible for optimal VNTR loci amplification. A modified PAGE system for VNTR separation was established. Under these conditions, new longer alleles in the 01580 locus were discovered and 025123 pattern changes in colorectal tumors were observed. These technical tips are valuable for detecting various amplified fragment length polymorphisms.

• Journal of Sasang Constitutional Medicine
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• v.9 no.2
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• pp.163-173
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• 1997

Amp-FLP is a one of the most frequently used human genetic analysis methods which adopts STR and VNTR typings. In this study, 100 genomic DNA samples of Taeum, Soyang and Soum constitution group were analysed by Amp-FLP method. The allele distribution of three STR loci(TH01, vWA and CSF1PO) and one VNTR locus(MCT118) of each different constitution group were investigated and the allele distribution was statistically evaluated. It was found out that the allele distribution of MCT118 and THO1 loci was not significantly different among different constitutions. However, the allele distribution of vWA showed p-value of 0.02056(Soyang group) and $2.41{\times}10^{-10}$(Soum group) which is much lower than significant level of p-value 0.05. Also, p-value of CSF1PO in Soum group was found out to be $4.65{\times}10^{-17}$. Therefore, it is expected that vWA and CSF1PO loci can be used as an indicator for gnenetic difference of different constiturion if the same result is obtained with sufficient number of samples.

• Tuberculosis and Respiratory Diseases
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• v.50 no.6
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• pp.668-675
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• 2001

Background : Non-smalll lung cancer(NSCLC) develops as a result of the accumulation of multiple genetic abnormalities. Loss of heterozygosity(LOH) is one of the most frequent genetic alterations that is found in NSCLC, and the chromosomal regions that display a high rate of LOH are thought to harbor tumor suppressor genes(TSGs). This study was done to determine the frequency of LOH in 21q with the aim of identifying potential TSG loci. Method : Thirty-nine surgically resected NSCLCs were analysed. Patients peripheral lymphocytes were used as the source of the normal DNA. Five microsatellite Inarkers of 21q were used to study LOH : 21q21.1(D21S1432, and D21S1994); 21q21.2-21.3(D21S1442) ; 21q22.1(21S1445) ; and 21q22.2-22.3(D21S266). The fractional allelic loss(FAL) in a tumor was calculated as the ratio of the number of markers showing LOH to the number of informative markers. Result : LOH for at least one locus was detected in 21 of 39 tumors(53.8%). Among the 21 tumors with LOH, 5(21.8%) showed LOH at almost all informative loci. Although statistically not significant, LOH was found more frequently in squamous cell carcinomas(15 of 23, 65.2%) than in adenocarcinomas(6 of 16, 37.5%). In the squamous cell carcinomas the frequency of LOH was higher in stage II-III (80.0%) than in stage I (53.8%). The FAL value in squamous cell carcinomas($0.431{\pm}0.375$) was significantly higher than that found in adenocarcinomas($0.l92{\pm}0.276$). Conclusion : These results suggest that LOH on 21q may be involved in the development of NSCLC, and that TSG(s) that contribute to the pathogenesis of NSCLC may exist on 21q.