• 제목/요약/키워드: STR genotyping

검색결과 13건 처리시간 0.02초

Choosing Optimal STR Markers for Quality Assurance of Distributed Biomaterials in Biobanking

  • Chung, Tae-Hoon;Lee, Hee-Jung;Lee, Mi-Hee;Jeon, Jae-Pil;Kim, Ki-Sang;Han, Bok-Ghee
    • Genomics & Informatics
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    • 제7권1호
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    • pp.32-37
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    • 2009
  • The quality assurance (QA) is of utmost importance in biobanks when archived biomaterials are distributed to biomedical researchers. For sample authentication and cross-contamination detection, the two fundamental elements of QA, STR genotyping is usually utilized. However, the incorporated number of STR markers is highly redundant for biobanking purposes, resulting in time and cost inefficiency. An index to measure the cross-contamination detection capability of an STR marker, the mixture probability (MP), was developed. MP as well as other forensic parameters for STR markers was validated using STR genotyping data on 2328 normal Koreans with the commercial AmpFlSTR kit. For Koreans, 7 STR marker (D2S1338, FGA, D18S51, D8S1179, D13S317, D21S11, vWA) set was sufficient to provide discrimination power of ${\sim}10^{-10}$ and cross-contamination detection probability of ${sim}1$. Interestingly, similar marker sets were obtained from African Americans, Caucasian Americans, and Hispanic Americans under the same level of discrimination power. Only a small subset of commonly used STR markers is sufficient for QA purposes in biobanks. A procedure for selecting optimal STR markers is outlined using STR genotyping results from normal Korean population.

PCR-based genotyping of Korean population for forensic applications

  • 류재송;구윤모;소재성
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.592-595
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    • 2000
  • In human chromosome, a short sequence of DNA has been repeated a number of times. These repeats are called variable number of tandem repeat(VNTR) or short tandem repeat(STR) which has short repeat core. VNTR and STR are used in the field of forensic science, evolution, and anthropology. In this work, we examined allele frequencies of 3 VNTR(YNZ22, NeuR, D21S11) and one STR(Humth01) in a Korean population sample by polymerase chain reaction(PCR) followed by high-resolution polyacrylamide gelelectrophoresis(PAGE) with silver staining. Subsequently, the polymorphism information content(PIC) was calculated : the highest PIC was observed for the NeuR locus(0.95680) and lowest for the Humth01 locus(0.75809).

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Optimization of a Multiplex DNA Amplification of Three Short Tandem Repeat Loci for Genetic Identification

  • Ryu, Jae-Song;Noh, Jae-Sang;Koo, Yoon-Mo;Lee, Choul-Gyun;So, Jae-Seong
    • Journal of Microbiology and Biotechnology
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    • 제10권6호
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    • pp.873-876
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    • 2000
  • Short tendem repeat (STR) loci have been used in the field of forensic science. There are literally hundreds of STR systems which have been mapped throughout the human genome. These STR loci are found in almost every chromosome in the genome. They may be amplified using a variety of PCR primers. In this study, a DNA genotyping system based on the multiplex amplification of highly polymorphic STR loci was developed. Three STR loci with nonoverlapping allele size ranges have been utilized in the multiplex amplification including the Neurotensin receptor gene, D21S11, and Human tyrosine hydroxylase gene. The optimal condition for triplex PCr was obtained in a solution with a total volume of $25{\mu}l$ containing 2.0 U of Taq polymerase, 3 mM of $MgCl_2$, $300{\mu}M$ of dNTP, 10 pmole of each primer set, an annealing temperature of $62^{\circ}C$, and 35 cycles. The optimized condition was successfully employed in a family paternity test.

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PCR Analysis of Four Length-Polmorphic Loci in Korea Population for Genotyping

  • Ryu, Jae-Song;Koo, Yoon-Mo;So, Jae-Seong
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권3호
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    • pp.169-173
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    • 2000
  • On human chromoscomes, a short sequence of DNA is known to repeat a number of times. These are called variable number of tandem repeat (VNTR) or short tandem respeat (STR) which has a short core. VNTR and STR are used in the filed of forensic science, evolution, and anthropology. In this work, we examined allele frequencies of one VNTR (YNZ22) and three STRs (NeuR, D21S11, Humth01) in a korean population sample by polymerase chain reaction (RCP) followed by high-resolution polyacrylamide gel electro-phoresis (PAGE) with silver stain. Subsequently, the polymorphism information content (PIC) was calculated : the hifhest PIC was observed in the NeuR locus (0.95680) and lowest in the Humth01 locus (0.75809).

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사상체질별 Short Tandem Repeat 대립유전자 빈도 (Short Tandem Repeat Allele Frequencies in Sasang Constitution)

  • 박화용;유현주;구임회;김종열
    • 사상체질의학회지
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    • 제21권1호
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    • pp.227-236
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    • 2009
  • 1. Objectives As a basic trial for identification of Sasang constitutional gene marker, we genotyped and analysed statistical relationships of STR(short tandem repeat) alleles and its distribution in each constitution. 2. Methods After obtaining basic constitutional data with questionnaire (QSCC II), decision of constitution was made by 3 different constitution specialists' diagnosis, and only the samples of specialists' agreement of each constitution by discussion were taken into this research. Using multiplex PCR kit, total 146 constitutional samples were amplified in 16 autosomal STR marker, genotyped, and analysed statistically. Among 16 markers, 15 were analysed in this study excluding the amelogenin marker is used for in gender identification. 3. Results and Conclusions It is difficult to determine the relationship between constitution and STR marker as the sample size is small, however, Penta D and vWA were shown to be related statistically with constitution. It has been know that STRs has no genetic informations, however there are some recent research results showing STRs as a regulatory element, relationship between microsatellite instability and repeat number and size, and post-transcriptional sigualing. STRs which is not known about its function currently, are proposed to have function and/or regulatory activities anyhow with Sasang constitution. It is believed that the results of this study can halp determine and deatify the markers related to Sasang Constitutional Medicien.

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Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • 한국동물위생학회지
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    • 제41권4호
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    • pp.257-262
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    • 2018
  • Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

유골에서 DNA 추출법 비교 연구: Ultrafiltration과 Column affinity (Evaluation of two DNA extraction methods on exhumed bone samples: Ultrafiltration versus column affinity)

  • 김순희;홍승범;브라이언 M. 켐프;박기원;한면수
    • 분석과학
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    • 제21권4호
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    • pp.338-343
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    • 2008
  • 발굴된 유골에서 DNA 추출은 DNA신원확인과정에서 매우 중요한 단계이지만 유골에 오염된 미생물 DNA와 사람 DNA가 동시에 추출되는 문제점이 있다. 이를 극복하기 위해 발굴된 10명의 유골 시료를 페놀-ultrafiltration 후 $QIAquick^{(R)}$ PCR Purification Kit (ultrafiltration법)와 $QIAamp^{(R)}$ DNA Mini kit(Column-affinity법)를 적용하여 전체(미생물+사람) DNA정량 및 사람 DNA정량 후 각각 STR마커를 분석하므로서 DNA 분리 효율성을 확인하였다. 회수된 전체 DNA양은 Column-affinity법($19.6ng/{\mu}L$)이 ultrafiltration법($16.0ng/{\mu}L$) 보다 1.2배 더 나은 결과를 보였으나 사람 DNA 정량 결과로는 Column-affinity 법($0.034ng/{\mu}L$) 보다 ultrafiltration법($0.498ng/{\mu}L$)이 14.6배 더 많은 회수율을 나타냈다. 유골에 있던 다량의 미생물 DNA가 사람 DNA와 섞여있을 경우, 사람 DNA가 실리카 친화성 컬럼에 부착되는 것이 미생물 DNA의 영향을 받으나, 필터의 경우는 미생물 DNA의 영향을 받지 않아서 ultrafiltration법이 높은 회수율을 보였다.

다중 중합효소연쇄반응을 이용한 핵산시료의 동정방법 (Simple Identification of DNA Samples Using Multiplex PCR)

  • 박화용;유현주
    • 동의생리병리학회지
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    • 제22권2호
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    • pp.427-430
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    • 2008
  • Serious controls and cares using ID numbers and barcode needed throughly to have appropriate management in clinical tissues and nucleic acids inventories because these samples are the most essential and important materials in the experimental research laboratories. While almost all of the laboratories using and handling DNA samples as starting materials in their research, problems such as mixing up of two or more different samples together, contamination with other samples, and/or mistakes can occur, especially when it comes with large number of samples. These problems are rather frequent even though researchers pay more attentions to be far away from these obstacles. It has been such a long time since PCR became useful as an important and essential biological research tool among lots of bio-scientific research methods. In this research, we tried to set up a simple and cost-effective genotyping method using PCR and agarose gels, instead of expensive automated machines, for identification and discrimination among those DNA samples, as a kind of low level quality control and sample inventory management.

총기 흔적흔에서의 low copy number(LCN) DNA 검출에 관한 연구 (Research on the detection of LCN DNA from traces on firearms)

  • 전충현;박성우
    • 분석과학
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    • 제24권1호
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    • pp.51-59
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    • 2011
  • 유전자 감식은 다양한 범죄현장에서 발견되는 생체시료의 분석을 통해 신원을 식별하는 중요한 법과학적 수사과정으로 자리 잡았다. 최근에는 범인이 사용했던 펜, 뺑소니 차량에서의 핸들, 기어, 각종 버튼스위치 등에 남겨져 있는 touch evidence-type sample로 알려져 있는 low copy number (LCN) DNA에서의 A-STR분석을 위해 의뢰되는 감정물들이 증가하는 추세에 있다. 본 연구에서는 총기의 뭉개진 지문 등에 남겨져 있는 touch evidence-type의 LCN DNA를 추출하고 유전자형의 분석 성공률을 확인하고 자 하였다. 4종류의 총기(M16, K1A, COLT 45 권총, M29 리볼버)를 각각 격발한 후 총기별로 4곳의 부위에서 시료를 채취한 다음 LCN DNA의 추출을 위해 Microkit과 $Prepfiler^{TM}$ 등 2종류의 시약을 이용하여 DNA 검출량과 유전자형 분석 성공률을 비교 분석하였다. 분석결과 $Prepfiler^{TM}$가 Microkit에 비해 평균 1.7배 DNA검출량이 많았으며, 유전자형 분석 성공률에 있어서도 Microkit은 0%인데 비해 $Prepfiler^{TM}$에서는 평균 24.9%의 성공률을 보였으며, K1A의 손잡이 부위에서 50.6%의 성공률을 나타냈다.