• Title/Summary/Keyword: STR genotyping

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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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    • v.7 no.1
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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

  • Ryu, Jae-Song;Gu, Yun-Mo;So, Jae-Seong
    • 한국생물공학회:학술대회논문집
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    • 2000.04a
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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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    • v.10 no.6
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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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    • v.5 no.3
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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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Short Tandem Repeat Allele Frequencies in Sasang Constitution (사상체질별 Short Tandem Repeat 대립유전자 빈도)

  • Park, Hwa-Yong;Yu, Hyun-Joo;Ku, Im-Hoi;Kim, Jong-Yeol
    • Journal of Sasang Constitutional Medicine
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    • v.21 no.1
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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
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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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.

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

  • Kim, Soonhee;Hong, Seungbeom;Kemp, Brian M.;Park, Kiwon;Han, Myunsoo
    • Analytical Science and Technology
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    • v.21 no.4
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    • pp.338-343
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    • 2008
  • Extraction of DNA from skeletal material is of great importance in the identification of human remains, but is particularly difficult because the high amount of microbial DNA was often co-extracted with human bone DNA. We found that a phenol/chloroform extraction, followed by ultrafiltration, and cleanup by via the $QIAquick^{(R)}$ PCR purification kit yields higher amounts of human genomic DNA compared with extraction by the column affinity $method^{(R)}$ alone. Ultrafiltration extraction of human DNA from ten exhumed bone samples yielded $0.041-1.120ng/{\mu}L$ DNA (mean = $0.498ng/{\mu}L$ DNA), and purification using the column affinity resulted in $0.016-0.064ng/{\mu}L$ DNA (mean = $0.034ng/{\mu}L$ DNA). Although the STR genotyping by the column affinity method was partially successful, all DNA samples by the ultrafiltration method produced full profiles from the multiplex PCR. The efficiency of STR genotyping was in accordance with the amounts of the human DNA extracted.

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

  • Park, Hwa-Yong;Yu, Hyun-Joo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.2
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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.

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

  • Jeon, Chung-Hyun;Park, Sung-Woo
    • Analytical Science and Technology
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    • v.24 no.1
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    • pp.51-59
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    • 2011
  • Genetic Identification has become an important forensic investigation method which discerns identity through analysis of physical samples discovered in various crime scenes. Recently more samples are being requested to undergo A-STR analysis of low copy number (LCN) DNA, which is known as touch evidence-type sample and left on various objects such as a pen briefly used by the criminal, the gear of the car used for driving, the handle, and various buttons inside a car. This research attempted to extract the LCN DNA of the touch evidencetype left on crushed fingerprints on firearms, etc. and examine the genotyping success rate. Four types of firearms (M16, K1A, COLT 45 Pistol, M29 Revolver) were fired individually and physical samples were gathered from four parts of each firearm. Subsequently, in order to extract the LCN DNA, Microkit and $Prepfiler^{TM}$ were used to compare and analyze the quantity of DNA extracted and the genotyping success rate. Analysis results showed that the quantity of DNA extracted by $Prepfiler^{TM}$ was on average 1.7 times higher than that of Microkit, and in genotype analysis success rate $Prepfiler^{TM}$ also demonstrated 24.9% on average in contrast to 0% for Microkit. In regards to the grip part of the K1A, $Prepfiler^{TM}$'s success rate was as high as 50.6%.