• Title/Summary/Keyword: DNA hybridization

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Species characterization of animal by DNA hybridization (DNA hybridization을 이용한 축종특이성 구명)

  • Lee, Myoung-heon;Kim, Sang-keun;Jung, Gab-soo;Park, Jong-myoung
    • Korean Journal of Veterinary Research
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    • v.39 no.3
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    • pp.513-522
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    • 1999
  • DNA hybridization assay using probes prepared from liver was carried out to identify species characterization of the domestic animals. Gel electrophoresis showed that the target DNA extracted from raw muscle were 1kb and uniform pattern while fragments size of heated muscle were irregular. Hybridization was performed by adding 200ng/ml probe in hybridization solution and incubating for 12 hours at $68^{\circ}C$. To obtain good discrimination, applied washing buffer and washing step differently depending on the species. The probes of pig, horse and dog formed the specific hybrids with each target DNA respectively. Although cross reaction was detected in cattle, goat and sheep but signal intensity among these species made the discrimination possible each other. Such pattern was the same in the cases of chicken, turkey and duck. The hybridization pattern of heated muscle was similar to that of raw muscle in general, but the signal intensity was inferior to that of raw muscle. Species identification between closely related animal species, hybridized using the target DNA of such closely related animal species as a blocking agent, remarkable increase of discrimination from the evident decrease of non specific reaction compared with the control group. In addition, in the admixture where certain meat was included in the beef, pork, chicken meat, we could find whether any unjust meat was admixed or not. In this case, detection limit of certain meat in admixture was 1%.

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SNP Detection of Arraye-type DNA Chip using Electrochemical Method (전기화학적 방법에 의한 신규 바이오칩의 SNP 검출)

  • 최용성;권영수;박대희
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.17 no.4
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    • pp.410-414
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    • 2004
  • High throughput analysis using a DNA chip microarray is powerful tool in the post genome era. Less labor-intensive and lower cost-performance is required. Thus, this paper aims to develop the multi-channel type label-free DNA chip and detect SNP (Single nucleotide polymorphisms). At first, we fabricated a high integrated type DNA chip array by lithography technology. Various probe DNAs were immobilized on the microelectrode array. We succeeded to discriminate of DNA hybridization between target DNA and mismatched DNA on microarray after immobilization of a various probe DNA and hybridization of label-free target DNA on the electrodes simultaneously. This method is based on redox of an electrochemical ligand.

Genomic DNA probe and purification of Theileria sergenti merozoites in Korean cattle (한우에 감염된 Theileria sergenti merozoite의 순수분리와 genomic DNA probe에 관한 연구)

  • Chae, Joon-seok;Lee, Joo-mook;Kwon, Oh-deog;Chae, Keon-sang
    • Korean Journal of Veterinary Research
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    • v.34 no.2
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    • pp.387-394
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    • 1994
  • To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from bovine erythrocytes. The infected erythrocytes were lysed by Aeromonas hydrophila(Ah-1) hemolysin, and the parasites were isolated by ultracentrifugation on a Percoll discontinuous density gradient. For construction of a T sergenti genomic DNA library, T sergenti DNA was digested with Pstl and the fragments were ligated into the PstI site of pUC19 before transformation of Escherichia coli JM83. Out of thousands of transformants obtained by transformation of E coli JM83 with the genomic library, three plasmids were chosen. The sizes of the inserted DNAs were 2.9kb(2.4kb and 0.5kb) in pKTS1, 4.3kb in pKTS2 and 1.5kb in pKTS3, respectively. The DNA fragments used as probe KTS1(2.4kb), KTS2(4.3kb) and KTS3(1.5kb) were labeled digoxigenin-11-dUTP for the Southern hybridization. In Southern hybridization, all of the probes(KTS1, KTS2 and KTS3) reacted specifically to T sergenti DNA, but not to bovine leucocyte DNA. In order to find out the sensitivities of the digoxigenin-11-dUTP-labeled KTS1 and KTS3 as the probes, purified merozoite DNA and bovine DNA (control) were checked by dot blot hybridization with the probes. Both of the probes, KTS1 and KTS3, detected as minimum amount of 975pg of the T sergenti DNA, but not bovine DNA even to 500ng.

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DNA fingerprinting patterns of 5 serotypes of Actinobacillus actinomycetemcomitans (Actinobacillus actinomycetemcomitans의 혈청형별 제한절편장 다변화에 관한 연구)

  • Choi, Jeom-Il;Koh, Myung-Yun;Yun, Il
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.365-375
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    • 1996
  • 5 serotypes(a, b, c, d, e) of Actinobacillus actinomycetemcomitans showed distinct hybridization patterns(DNA fingerprinting patterns) when the bacterial DNA were hybridized with randomly cloned 4.7-Kb sized DNA probe. The sizes of hybridized bands in each serotypes were different among serotypes and represented unique patterns of hybridization with the probe used. The serotype a showed two bands of fingerprinting patterns: 23.1 kb and 2.5 kb respectively. Serotype b and c showed single band: 6.6 kb and 9.5 kb, respectively. Serotype d and e showed two bands of hybridization: 23.1 kb and 2.8 kb, and 23.7 kb and 2.1 kb, respectively. The results indicate that this standard fingerpriting patterns of DNA hybridization with 4.7 kb probe can be further used for genotyping clinical isolates of Actinobacillus 8ctinomycetemcomitansand its relevance with periodontal disease activity.

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Design of Temperature Regulation for DNA Kernel to Satisfy Positive Definiteness (DNA 커널이 양한정 조건을 만족시키기 위한 온도 조절 디자인)

  • Noh, Yung-Kyun;Kim, Cheong-Tak;Zhang, Byoung-Tak
    • Proceedings of the Korean Information Science Society Conference
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    • 2007.06b
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    • pp.15-20
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    • 2007
  • 기존의 연구는 DNA 커널을 통한 기계 학습이 DNA 분자들을 통한 in vitro 실험을 통해 가능함을 보였다. 이 때, DNA 커널을 통한 분류 분제는 온도 조절을 통해 양한정(positive definite) 조건을 만족시킬 때 분류 문제를 잘 풀며, 양한정 조건을 만족시키기 위한 조건으로 높은 온도에서 시작하여 온도를 내리며 hybridization시키는 방법을 제안하였다. 이 논문에서는 보다 정량적인 분석을 통해서 이 hybridization 방법이 양한정 조건을 만족시키기에 적합한 방법임을 보이고, 간단한 hybridization 모델을 통해 양한정 조건을 만족시킬 수 있는 hybridization 온도 계획의 충분 조건을 유도한다. 또한 시작 온도와 끝 온도의 경계 조건으로 제시되는 이 충분 조건을 통해 현실적인 온도 조절 계획을 위한 시퀀스의 코딩 방법을 알게 된다.

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Detection of SNP Using Microelectrode Array Biochip (마이크로전극어레이형 바이오칩을 이용한 SNP의 검출)

  • Choi, Yong-Sung;Kwon, Young-Soo;Paek, Dae-Hee
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2004.07b
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    • pp.845-848
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    • 2004
  • High throughput analysis using a DNA chip microarray is powerful tool in the post genome era. Less labor-intensive and lower cost-performance is required. Thus, this paper aims to develop the multi-channel type label-free DNA chip and detect SNP (Single nucleotide polymorphisms). At first, we fabricated a high integrated type DNA chip array by lithography technology. Various probe DNAs were immobilized on the microelectrode array. We succeeded to discriminate of DNA hybridization between target DNA and mismatched DNA on microarray after immobilization of a various probe DNA and hybridization of label-free target DNA on the electrodes simultaneously. This method is based on redox of an electrochemical ligand.

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생물 공학 의약품의 품질관리에 관한 연구(비방사능 물질 표지법을 이용한 숙주유래 DNA의 검출법 개발)

  • 용군호;민홍기;김창민;오호정;한강현;최규실
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.59-59
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    • 1993
  • 비방사능물질인 Biotin을 DNA probe에 표지하여 nonisotopic hybridization 방법을 사용하여 감도를 높임으로써, 손쉽게 생물공학 의약품의 품질관리에 사용되도록 하였다. Bethesda Research Laboratories(BRL) 회사 제품인 biotinylated probes-avidin alkaline phosphatase를 이용한 chemiluminescene detection방법으로 행하여 λ phage DNA, yeast DNA, E.coil DNA의 한계 검출 농도를 알아내고, 생물 공학 제품에 적용하였다. Dot blot hybridization 방법으로 행하여 λ phage DNA는 0.1pg, Yeast DHA는 4.5pg, E. coli DNA는 8.9pg까지 검출되었고, 기존 생물공학 제품에서는 숙주 유래 DNA가 전혀 검출되지 않았다.

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Improvement in Sensitivity by Increasing the Frequency of SAW Sensors for DNA Detection (DNA 측정용 SAW 센서의 주파수 증대에 의한 감도향상)

  • Sakong, Jung-Yul;Kim, Jae-Ho;Lee, Soo-Suk;Roh, Yong-Rae
    • The Journal of the Acoustical Society of Korea
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    • v.26 no.1
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    • pp.42-47
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    • 2007
  • In this paper. we have studied improvement in sensitivity by increasing the frequency of SAW sensors for detecting the immobilization and hybridization of DNA. The sensor consists of twin SAW delay lines operating at 200MHz, a sensing channel and a reference channel. fabricated on $36^{\circ}$ rotated Y-cut X-propagation $LiTaO_3$ crystals. The optimum concentration of probe and target DNA was decided for the improvement of detection mechanism. and digital syringe pump system was used to reduce the human errors. The hybridization between immobilized probe DNA and target DNA on the gold-coated delay line results in mass loading on the delay line of the sensing channel. Thus, the relative frequency change was monitored in relation to the mass loading. The measurement results showed a good response of the sensor to the DNA hybridization with a maximum sensitivity level up to 0.066ng/m1/Hz.

Detection of DNA Hybridization Characteristics Using Electrochemical methods (전기화학법을 이용한 DNA Hybridization 특성 검출)

  • Kim, Do-Kyun;Chang, Jeong-Soo;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2002.07c
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    • pp.1569-1571
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    • 2002
  • The determination of DNA hybridization can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, The determination of hybridization is very important for the improvement of DNA detection system. In this study, we report the characterization of the DNA hybridization by the electricalchemical methods. A new electrochemical biosensor is described for voltammetric detection of gene sequence related to probe oligonucleotide of bacterium Escherichia coli O157:H7. The biosensor involves the immobilization of a 18-mer probe oligonucleotide, which is complemetary to a specific gene sequence related to Escherichia coli O157:H7 on a gold electrode through specific adsorption. The probe oligonucleotide was used to determine the amount of target oligonucleotide in solution using mitoxantrone(MTX) as the electrochemical indicators. The cathodic peak currents $(I_{peak})$ of MTX were linearly related to the concentration of the target oligonucleotide sequence in the range $1[{\mu}M]{\sim}0.1[nM]$. The detection limit of this approach was 0.01[nM]. In addition, these indicators were capable of selectivity discriminating against various mismatching condition.

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