• 제목/요약/키워드: polymorphism detection

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Single-Strand Conformation Polymorphism Analysis by Microchip Electrophoresis for the Rapid Detection of Point Mutation in Human Obesity Gene

  • Kang, Seong-Ho;Jang, Soo-Young;Park, Sang-Kyu
    • Bulletin of the Korean Chemical Society
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    • 제27권9호
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    • pp.1346-1352
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    • 2006
  • We describe an effective method of microchip electrophoresis (ME) based on single strand conformation poly-morphism (SSCP) analysis to rapidly detect the point mutation, Leu72Met, in a human obesity gene. The 207-bp dsDNA in the Leu72Met region, an estimate of the child obesity DNA mutant, was amplified by polymerase chain reaction (PCR) and submitted to a conventional glass microchip analysis with a sieving matrix of 1.75% poly(vinylpyrrolidone) (Mr 1 300 000), 1.0% poly(ethyleneoxide) (Mr 600 000) and 5.0% w/w glycerol. When combined with base stacking (BS) with hydroxide ions, the SSCP-ME provided rapid analysis as well as sensitive detection. The detection sensitivity was effectively enhanced in the OH- concentration range of 0.01-0.025 M NaOH. The sensitivity and speed of this ME-based SSCP methodology for the rapid detection of Leu72Met point mutations makes this an attractive method for diagnosing childhood obesity in a clinical diagnostic laboratory.

비수식화 DNA를 이용한 SNP의 검출 (SNP (Single Nucleotide Polymorphism) Detection Using Indicator-free DNA)

  • 최용성;박대희;권영수
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2003년도 추계학술대회 논문집 Vol.16
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    • pp.224-226
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    • 2003
  • In this paper, we succeeded SNP discrimination of DNA hybridization on microarray using new electrochemical system. Using the electrochemical method with a label-free DNA has Performed DNA chip microarray. This method is based on redox of an electrochemical ligand. We developed scanning system with high performance.

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AFLP marker를 이용한 콩의 유전적 다양성과 유전분리 분석 (Diversity and Inheritance of AFLP Markers in Wild and Cultivated Soybeans)

  • 김용호;윤홍태
    • 한국자원식물학회지
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    • 제17권3호
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    • pp.265-271
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    • 2004
  • AFLP marker의 유용성 을 알아보고자 재배콩과 야생콩을 대상으로 유전적 다양성과 유전분리 현상을 분석하였다. 공시 재료들의 polymorphism은 재배 콩과 야생 콩에서 각각 평 균 2 9%와 12.2%의 polymorphism을 보였으며, 재배 콩과 야생 콩에서 공히 유전적 다양성을 보인 DNA단편은 11개 primer 평균 24개를 나타내었다. Primer 조합별로도 polymorphism에 다양한 차이가 있었는데 평균 22.9%로 13.0-38.5%의 변이를 나타내었다. 재배 콩 간의 교잡후대(화엄풋콩 ${\times}$ PI417479) F$_2$집단에서 AFLP marker의 유전분리 양상을 분석한 결과 3 : 1의 분리 비를 따르는 것으로 판단되었다.

Evaluation of the Frequency of the IL-28 Polymorphism (rs8099917) in Patients with Chronic Hepatitis C Using Zip Nucleic Acid Probes, Kerman, Southeast of Iran

  • Iranmanesh, Zahra;Mollaie, Hamid Reza;Arabzadeh, Seyed Alimohammad;Zahedi, Mohammad Javad;Fazlalipour, Mehdi;Ebrahimi, Saeede
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권5호
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    • pp.1919-1924
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    • 2015
  • Polymorphisms in the region of the interleukin IL-28 gene on chromosome 19 have been related with clearance of hepatitis C virus (HCV), a major human pathogen responsible for chronic hepatitis, cirrhosis and hepatocellular carcinoma. About 3% of the world's population is infected with HCV. The long-term response to therapy is influenced by many host and viral factors, and recent evidence has indicated that some host genetic polymorphisms related to IL-28 are the most powerful predictors of virological response in patients with HCV. This study assessed frequency of the IL-28 polymorphism (rs8099917) in 50 patients (39 men and 11 women) with chronic hepatitis C using ZNA probe real time PCR new method. All patients were tested for genotype of HCV and the HCV viral load. In parallel, the levels of SGOT, SGPT and ALK enzymes were assessed. Treatment using Peg-interferon alpha with ribavirin was conducted for patients and subsequently samples were collected to detect any change in viral load or liver enzyme rates. The overall frequency of the TT allele is 74%, TG allele 20% and GG allele 6% and the percent of patients who had T allele was 84%. Clear reduction in viral load and liver enzymes was reported in patients with the T allele. Especially for genotype 1 which is relatively resistant to treatment, these alleles may have a role in this decline. In conclusion, we showed that IL-28 polymorphism rs8099917 strongly predicts virological response in HCV infection and that real-time PCR with Zip nucleic acid probes is a sensitive, specific and rapid detection method for detection of SNPs which will be essential for monitoring patients undergoing antiviral therapy.

한우 혈액에서 PCR을 이용한 Mycobacterium avium ssp paratuberculosis의 검출 (Detection of Mycobacterium avium ssp paratuberculosis in Korean Cattle by the Polymerase Chain Reaction)

  • 김광현;곽길한;송희종;조정곤
    • 한국임상수의학회지
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    • 제27권1호
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    • pp.23-28
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    • 2010
  • Mycobacterium avium ssp paratuberculosis, intracellular bacteria that can cause chronic granulomatous enteritis in cattle, continues to pose significant economic losses and health problem with high prevalence. The purpose of this study is the polymerase chain reaction (PCR)-base strategy for early detection of M. avium ssp paratuberculosis in whole blood. Blood samples were collected from korean cattles in Jeonbuk, Korea. The 16 out of 88 serum samples were detected M. partuberculosis by ELISA. Then samples of infected 8 Korean cattles were amplified by PCR. The PCR amplified targets are 16s rDNA and heat shock protein 65kDa (hsp 65). The 16s rDNA provided a highly sensitive and specific tool for the direct detection of mycobacteria. In addition M. avium was confirmed characteristically by the hsp65. Finally there were sure to M. avium ssp paratuberculosis by IS900 PCR. The restriction fragment length polymorphism was identified by PCR amplifications and subsequence restriction enzyme digestions with Pst I of a hsp65. These results indicate that confirm M. avium with 16s rDNA, hsp65 and a restriction fragment length polymorphism in the hsp65 gene can be seem the other pattern. Therefore, these results can be used for clinical direct detections of M. avium ssp paratuberculosis in whole blood of Korean cattle and also to be used epidemiological researches.

The Application of RNA Transcript Conformation Polymorphism in Resolving Mixed Infection of PVY Isolates

  • Maslenin, Ludmila;Rosner, Arie
    • The Plant Pathology Journal
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    • 제20권4호
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    • pp.308-312
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    • 2004
  • A method based on RNA-transcript conformation polymorphism (TCP) was tested for detection of two PVY isolates in a mixed infection. Differences in electrophoretic mobility of RNA transcripts copied from PCR products of each virus isolate enabled the distinction between the two virus isolates in a mixed infection. The identities of the RNA transcripts and hence of the infecting virus isolates were determined by annealing to reference oligonucleotides containing unique strain-specific sequences visualized by retardation of transcript mobility in gel. The ratio at which both virus isolates could be detected was as low as 1:10. The suitability of this procedure for the study of mixed virus infections is discussed.

SNP-Based Fetal DNA Detection in Maternal Serum Using the HID-Ion AmpliSeqTM Identity Panel

  • Cho, Sohee;Lee, Ji Hyun;Kim, Chong Jai;Kim, Moon Young;Kim, Kun Woo;Hwang, Doyeong;Lee, Soong Deok
    • The Korean Journal of Legal Medicine
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    • 제41권2호
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    • pp.41-45
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    • 2017
  • Fetal DNA (fDNA) detection in maternal serum is a challenge due to low copy number and the smaller size of fDNA fragments compared to DNA fragments derived from the mother. Massively parallel sequencing (MPS) is a useful technique for fetal genetic analysis that is able to detect and quantify small amounts of DNA. In this study, seven clinical samples of maternal serum potentially containing fDNA were analyzed with a commercial single nucleotide polymorphism (SNP) panel, the HID-Ion $AmpliSeq^{TM}$ Identity Panel, and the results were compared to those from previous studies. Reference profiles for mothers and fetuses were not available, but multiple Y chromosomal SNPs were detected in two samples, indicating that fDNA was present in the serum and thereby validating observations of autosomal SNPs. This suggests that SNP-based MPS can be valuable for fDNA detection, thereby offering an insight into fetal genetic status. This technology could also be used to detect small amounts of DNA in mixed DNA samples for forensic applications.

Detection of Polymorphism of Growth Hormone Gene for the Analysis of Relationship between Allele Type and Growth Traits in Karan Fries Cattle

  • Pal, Aruna;Chakravarty, A.K.;Bhattacharya, T.K.;Joshi, B.K.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권10호
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    • pp.1334-1337
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    • 2004
  • The present study was conducted to detect polymorphism at growth hormone gene in Karan Fries bulls. A 428 bp fragment of growth hormone gene spanning over $4^{th}$exon, $4^{th}$intron and $5^{th}$ exon was amplified and digested with AluI restriction enzyme to identify polymorphism at this locus. Karan Fries bulls were found to be polymorphic at this locus. Two genotypes LL and LV were identified in Karan Fries with higher allelic frequency for L allele. In Karan Fries males, the average birth weight, 3 months body weight and daily body weight gains of LL homozygotes were significantly higher than that of LV heterozygotes. Genetic distances of KF bulls with respect to genotype along with 3 months body weight and average daily body weight gain forms a single cluster of bulls with LL genotype, while individuals with LV genotype forms three distinct clusters indicating more influence of L allele on growth traits.

Comparison of Normalization Methods for Defining Copy Number Variation Using Whole-genome SNP Genotyping Data

  • Kim, Ji-Hong;Yim, Seon-Hee;Jeong, Yong-Bok;Jung, Seong-Hyun;Xu, Hai-Dong;Shin, Seung-Hun;Chung, Yeun-Jun
    • Genomics & Informatics
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    • 제6권4호
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    • pp.231-234
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    • 2008
  • Precise and reliable identification of CNV is still important to fully understand the effect of CNV on genetic diversity and background of complex diseases. SNP marker has been used frequently to detect CNVs, but the analysis of SNP chip data for identifying CNV has not been well established. We compared various normalization methods for CNV analysis and suggest optimal normalization procedure for reliable CNV call. Four normal Koreans and NA10851 HapMap male samples were genotyped using Affymetrix Genome-Wide Human SNP array 5.0. We evaluated the effect of median and quantile normalization to find the optimal normalization for CNV detection based on SNP array data. We also explored the effect of Robust Multichip Average (RMA) background correction for each normalization process. In total, the following 4 combinations of normalization were tried: 1) Median normalization without RMA background correction, 2) Quantile normalization without RMA background correction, 3) Median normalization with RMA background correction, and 4) Quantile normalization with RMA background correction. CNV was called using SW-ARRAY algorithm. We applied 4 different combinations of normalization and compared the effect using intensity ratio profile, box plot, and MA plot. When we applied median and quantile normalizations without RMA background correction, both methods showed similar normalization effect and the final CNV calls were also similar in terms of number and size. In both median and quantile normalizations, RMA backgroundcorrection resulted in widening the range of intensity ratio distribution, which may suggest that RMA background correction may help to detect more CNVs compared to no correction.