• Title/Summary/Keyword: DNA genotyping

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Preprocessing technology on DNA genotyping for personal verification (개인인식을 위한 DNA Genotyping의 전처리 기법)

  • 임효빈;오옥균;공은배
    • Proceedings of the Korean Information Science Society Conference
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    • 2004.04b
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    • pp.586-588
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    • 2004
  • 최근 DNA 관련 기술의 개발이 활발하게 이루어지고 있고, 그에 따라 DNA를 개인인식에 사용마고자 하는 시도가 실시간 이용가능성이 높은 DNA 칩 기술과 합쳐져 매우 중요한 이슈로 떠오르고 있다. DNA를 분석하기 위해서는 생물학적 실험이 필수적으로 따르게 되는데 이러한 실험 결과를 인식에 적용하기 위해서는 적절한 전처리가 필요하다. 본 논문에서는 여러 장점들로 인해 최근 DNA분석 기술로 주목받고 있는 모세관 전기영동법을 사용하여 DNA를 분석하고, 그 분석물을 개인인식을 위해 genotyping하는 과정에서 전처리가 요구되는 각 경우들에 대해 논하고 적절한 필터링 기법들을 제시한다.

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DNA Pooling as a Tool for Case-Control Association Studies of Complex Traits

  • Ahn, Chul;King, Terri M.;Lee, Kyusang;Kang, Seung-Ho
    • Genomics & Informatics
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    • v.3 no.1
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    • pp.1-7
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    • 2005
  • Case-control studies are widely used for disease gene mapping using individual genotyping data. However, analyses of large samples are often impractical due to the expense of individual genotyping. The use of DNA pooling can significantly reduce the number of genotyping reactions required; hence reducing the cost of large-scale case-control association studies. Here, we discuss the design and analysis of DNA pooling genetic association studies.

Electrochemical measurement for analysis of DNA sequence (DNA 염기서열 분석을 위한 전기 화학적 측정법)

  • Jo, Seong-Bo;Hong, Jin-Seop;Kim, Yeong-Mi;Park, Jeong-Ho
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.51 no.2
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    • pp.92-97
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    • 2002
  • One of the important roles of a DNA chip is the capability of detecting genetic diseases and mutations by analyzing DNA sequence. For a successful electrochemical genotyping, several aspects should be considered including the chemical treatment of electrode surface, DNA immobilization on electrode, hybridization, choice of an intercalator to be selectively bound to double standee DNA, and an equipment for detecting and analyzing the output signal. Au was used as the electrode material, 2-mercaptoethanol was used for linking DNA to Au electrode, and methylene blue was used as an indicator that can be bound to a double stranded DNA selectively. From the analysis of reductive current of this indicator that was bound to a double stranded DNA on an electrode, a normal double stranded DNA was able to be distinguished from a single stranded DNA in just a few seconds. Also, it was found that the peak reduction current of indicator is proportional to the concentration of target DNA to be hybridized with probe DNA. Therefore, it is possible to realize a sim71e and cheats DNA sensor using the electrochemical measurement for genotyping.

Method Discrimination for Product Traceability and Identification of Korean Native Chicken using Microsatellite DNA (초위성체를 이용한 한국 재래닭의 원산지 추적 및 개체 식별 방법에 관한 연구)

  • Park, Mi-Hyun;Oh, Jae-Don;Jeon, Gwang-Joo;Kong, Hong-Sik;Sang, Byong-Don;Choi, Chull-Hwan;Yeon, Sung-Hum;Cho, Byong-Wok;Lee, Hak-Kyu
    • Korean Journal of Organic Agriculture
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    • v.12 no.4
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    • pp.451-461
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    • 2004
  • In an animals, identification system has been widely used by ear tag with dummy code and blood typing for parernity. Also, genotyping methods were using for useful mean of individual identification for live animals. In the case of genotyping estimation of gene in population of korean native chicken. In this study, we tested for development of genetic markers used it possible to determination of individual identification system. The candidate genetic markers were used already bow 10 of microstalite DNA sequence information in chromosome No. 1 and 14. Result of analysis for genotyping, the number of alleles of those microstatelites DNA was shown minimal 3 to 12 and the heterozygote expression frequency range was shown from 0.617 to 0.862. In our result, effective number of allele for each microsatellites DNA was shown 3~7, and the accuracy of individual identification was shown nearly 100%, when used with 6 genetic marker. This study was about genotyping method for identification used specific genetic marker form microsatellite DNA in the brand marketing of korean native chicken. Our results suggest that genotyping method used specific genetic marker from microsatellite DNA might be very useful for determination of individual identification.

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Noninvasive fetal RHD genotyping using cell-free fetal DNA incorporating fetal RASSF1A marker in RhD-negative pregnant women in Korea

  • Han, Sung-Hee;Yang, Young-Ho;Ryu, Jae-Song;Kim, Young-Jin;Lee, Kyoung-Ryul
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.100-108
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    • 2015
  • Purpose: Conventional methods for the prenatal detection of fetal RhD status involve invasive procedures such as fetal blood sampling and amniocentesis. The identification of cell-free fetal DNA (cffDNA) in maternal plasma creates the possibility of determining fetal RhD status by analyzing maternal plasma DNA. However, some technical problems still exist, especially the lack of a positive control marker for the presence of fetal DNA. Therefore, we assessed the feasibility and accuracy of fetal RHD genotyping incorporating the RASSF1A epigenetic fetal DNA marker from cffDNA in the maternal plasma of RhD-negative pregnant women in Korea. Materials and Methods: We analyzed maternal plasma from 41 pregnant women identified as RhD-negative by serological testing. Multiplex real-time PCR was performed by amplifying RHD exons 5 and 7 and the SRY gene, with RASSF1A being used as a gender-independent fetal epigenetic marker. The results were compared with those obtained by postnatal serological analysis of cord blood and gender identification. Results: Among the 41 fetuses, 37 were RhD-positive and 4 were RhD-negative according to the serological analysis of cord blood. There was 100% concordance between fetal RHD genotyping and serological cord blood results. Detection of the RASSF1A gene verified the presence of cffDNA, and the fetal SRY status was correctly detected in all 41 cases. Conclusion: Noninvasive fetal RHD genotyping with cffDNA incorporating RASSF1A is a feasible, reliable, and accurate method of determining fetal RhD status. It is an alternative to amniocentesis for the management of RhD-negative women and reduces the need for unnecessary RhIG prophylaxis.

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.

Strain-specific Detection of Bacillus Anthracis using Multiple-locus Variable-number Tandem Repeat Analysis (Multiple-locus Variable-number Tandem Repeat 분석을 사용한 Bacillus Anthracis 균주간 특이성 규명)

  • Jung, Kyoung-Hwa;Kim, Sang-Hoon;Kim, Seong-Joo;Kim, Ji-Cheon;Chai, Young-Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.14 no.2
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    • pp.305-312
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    • 2011
  • Bacillus anthracis(Ba) is a Gram-positive spore-forming bacterium that causes the disease anthrax. The feature of Ba is the presence of two large virulence plasmids, pXO1 and pXO2. Molecular genotyping of Ba has been difficult to the lack of polymorphic DNA marker. Ba isolated from Korea has been genotyped using various nucleotide analysis methods, such as 16s rDNA sequencing and multiple-locus variable-number tandem repeat (MLVA) analysis. We identified genotypes that represent a genetic lineage in the B1 cluster. This study emphasized the need to perform molecular genotyping when attempting to verify a strain-specific Ba.

Molecular Typing of Vibrio parahaemolyticus by Repetitive Element-PCR (rep-PCR) (Repetitive Element-PCR (rep-PCR)을 이용한 Vibrio parahaemolyticus 의 분자유전학적 아형 분류)

  • Kim, Won Sik;Hong, Seung Bok;Lee, Kyung;Lee, Jung Nam;Shin, Kyeong Seob
    • Korean Journal of Clinical Laboratory Science
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    • v.36 no.1
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    • pp.1-6
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    • 2004
  • The enterobacterial repetitive intergenic consensus (ERIC)-PCR is a recently described DNA fingerprinting technique based on amplification of repetitive element distributed in bacteria. We applied of ERIC-PCR to clinical isolates of Vibrio parahaemolyticus and other bacteria associated diarrhea. Twenty isolates of V. parahaemolyticus were used for intragenic genotyping, which were isolated from 2001 to 2002 in Chungbuk National University hospital. For interspecies genotyping, V. vulnificus, V. alginolyticus, V. parahaemolyticus, Escherichia coli, Salmonella and Shigella spp. were used. The genotyping were analyzed by ERIC-PCR. The genotyping of V. parahaemolyticus were grouped two major pattern (A, B) and were subdivided into 10 subtypes (A1, A2, B1-B8) by ERIC-PCR. These method distinctly differentiated bacterial species associated diarrhea. Those results show that ERIC-PCR can be reliable and efficient method for genotyping of V. parahaemolyticus and bacteria associated diarrhea.

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Diet Folate, DNA Methylation and Polymorphisms in Methylenetetrahydrofolate Reductase in Association with the Susceptibility to Gastric Cancer

  • Gao, Shang;Ding, Li-Hong;Wang, Jian-Wei;Li, Cun-Bao;Wang, Zhao-Yang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.299-302
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    • 2013
  • Methylenetetrahydrofolate reductase (MTHFR) has been reported to be associated with DNA methylation, an epigenetic feature frequently found in gastric cancer. We conducted a case-control study to explore the association of MTHFR C677T polymorphisms with gastric cancer risk and its relation with the DNA methylation of COX-2, MGMT, and hMLH1 genes. Genotyping of P16, MGMT and HMLH1 was determined by methylation-specific PCR after sodium bisulfate modification of DNA, and genotyping of MTHFR C677T was conducted by TaqMan assays using the ABI Prism 7911HT Sequence Detection System. Folate intake was calculated with the aid of a questionnaire. Compared with the MTHFR 677CC genotype, the TT genotype was significantly associated with 2.08 fold risk of gastric cancer when adjusting for potential risk factors. Individuals who had an intake of folate above $310{\mu}g$/day showed protective effects against gastric cancer risk. The effect of MTHFR C677T polymorphisms on the risk of gastric cancer was modified by folate intake and methylation status of MGMT (P for interaction <0.05).

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%.