• 제목/요약/키워드: DNA probes

검색결과 245건 처리시간 0.03초

DNA Chip 제작을 위한 Microarrayer의 개발 (Development of Microarrayer for DNA Chips)

  • 김석열;정남수;이재성;김상봉
    • 대한기계학회:학술대회논문집
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    • 대한기계학회 2003년도 춘계학술대회
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    • pp.899-904
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    • 2003
  • Microarrayer makes DNA chip and microarray that contain hundreds to thousands of immobilized DNA probes on surface of a microscope slide. This paper shows the development results for a printing type of microarrayer. It realizes a typical, low-cost and efficient microarrayer for generating low density microarray. The microarrayer is developed by using a robot of three-axes perpendicular type. It is composed of a computer-controlled three-axes robot and a pen tip assembly. The key component of the arrayer is the print-head containing the tips to immobilize cDNA, genomic DNA or similar biological material on glass surface. The robot is designed to automatically collect probes from two 96-well plates with up to 32 tips at the same time. To prove the performance of the developed microarrayer, the general water types of inks such as black, blue and red. The inks are distributed at proper positions of 96 well plates and the three color inks are immobilized on the slide glass under the operation procedure. As the result of the test, it can be shown that it has sufficient performance for the production of low integrated DNA chip consisted of 96 spots within 1 $cm^2$ area.

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DNA Ligand - Redox Active Molecule Conjugates as an Electrochemical DNA Probe

  • Ihara, Toshihiro;Maruo, Voshiyuki;Uto, Yoshihiro;Takenaka, Shigeori;Takagi, Makoto
    • 분석과학
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    • 제8권4호
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    • pp.887-894
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    • 1995
  • Toward the development of universal, sensitive, and convenient method of DNA (or RNA) detection, two kinds of electrochemically active DNA ligands. acridine - viologen and oligonucleotide - ferrocene conjugate, were prepared. Thermodynamic and electrochemical study revealed that these probes bound strongly to DNA, and showed a typical cyclic voltammograms, indicating a potential for use as a reversible electrochemical labelling agent for DNA. Especially, using the electrochemically active oligonucleotide, we have been able to demonstrate the detection of DNA at femtomole levels by HPLC equipped with ordinary electrochemical detector (ECD). These results lead to the conclusion that the redox-active probes are very useful for the microanalysis of nucleic acid due to the stabilily of the complexes, high detection sensitivity, and wide applicability to the target structures (single- and double strands) and sequences.

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Detection of Prevotella intermedia and Prevotella nigrescens using Pn17 and Pn34 DNA Probes

  • Park, Chan-Ho;Kim, Pan-Soon;Kim, Hwa-Sook;Min, Jeong-Bum;Hwang, Ho-Keel;Jang, Hyun-Sun;Cho, Ki-Woon;Baek, Dong-Heon;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제35권1호
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    • pp.13-19
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    • 2010
  • The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.

재조합 DNA probe에 의한 Fusarium oxysporum 분화형간의 분류 및 유전적 변이 분석 (Classification and Genetic Variation Analysis Among Formae Speciales of Fusarium oxysporum by Using Recombinant DNA Probes)

  • 김영태;김홍기
    • 한국균학회지
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    • 제25권4호통권83호
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    • pp.362-368
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    • 1997
  • 재조합 DNA probe를 이용하여 국내 Fusarium oxysporum 분화형간의 분류 가능성을 탐색하고 그들의 유전적 변이를 분석하였다. F. oxysporum f. sp. lycopersici, cucumerinum, fragariae, garlic, sesami 등 5종의 분화형을 공시하여 RFLP 분석을 실시하였다. Repetitive copy clone인 세종의 재조합 클론 pFC46, pFC52, pFC57을 이용하여 HindIII로 처리한 F. oxysporum의 genomic DNA에 대해 southern hybridization한 결과 나타난 밴드의 양상은 분화형에 따라 차이가 명확히 밝혀져 이들을 이용한 분류가 가능하였다. 또한 RFLP 분석 결과 f. sp. sesami는 다른 분화형에 비해 다소 변이가 심했으나 다른 분화형들은 변이가 거의 없어 f. sp. sesami를 제외한 f. sp. lycopersici 등 4종의 Fusarium oxysporum 분화형의 균주들은 채집 지역에 관계없이 대체로 유전적으로 안정되어 있는 것으로 판단되었다. Hybridization밴드의 양상에 기초하여 유전적 유연관계를 집괴 분석한 결과 각 분화형별로 유사도가 매우 높게 별도의 유사군을 형성하였다.

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Indicator-free DNA Chip Array Using an Electrochemical System

  • Park, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • 제4C권4호
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    • pp.133-136
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    • 2004
  • This research aims to develop a DNA chip array without an indicator. We fabricated a microelectrode array through photolithography technology. Several DNA probes were immobilized on an electrode. Then, target DNA was hybridized and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between the DNA probe and mismatched DNA in an anodic peak. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

Application of heterogeneous RNA probes labeled immuno-fluorescent agent DIG for the screening a noble gene in cucumber

  • Kim, Dai-Hee;Kim, Byung-Oh;Park, Gyu-Hwan;Takahashi, Hideyuki;Kim, Kyung-Min
    • Plant Resources
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    • 제7권2호
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    • pp.116-122
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    • 2004
  • We tried to isolating a noble gene from cucumber library with heterogeneouse RNA probe labeled DIG of Arabidopsis PIN3 gene. Two kinds of RNA probes which had no significant homology each others, were designed from the 5'- and 3'- prime nucleotides of the AtPIN3 gene. In the first and second screenings of the cDNA library of cucumber with the probes, two positive clones were identified with specific duplicate signals. However, we isolated cDNA fragments homologous with putative nucleases from Nicotiana, Arabidopsis, Cordialis, and Oryza sativa, there was no significant homology with any other PIN family genes.

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Development of a Microarrayer for DNA Chips

  • Kim Sang Bong;Jeong Nam Soo;Kim Suk Yeol;Lee Myung Suk
    • Fisheries and Aquatic Sciences
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    • 제5권1호
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    • pp.36-42
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    • 2002
  • Microarrayer is used to make DNA chip and microarray that contain hundreds to thousands of immobilized DNA probes on surface of a microscope slide. This paper shows the develop-ment results for a printing type of microarrayer. It realizes a typical, low-cost and efficient microarrayer for generating low density micro array. The microarrayer is developed by using a prependicular type robot with three axes. It is composed of a computer-controlled three-axes robot and a pen tip assembly. The key component of the arrayer is the print-head containing the tips to immobilize cDNA, genomic DNA or similar biological material on glass surface. The robot is designed to automatically collect probes from two 96-well plates with up to 12 pens at the same time. To prove the performance of the developed microarrayer, we use the general water types of inks such as black, blue and red. The inks are distributed at proper positions of 96 well plates and the three color inks are immobilized on the slide glass under the operation procedure. As the result of the test, we can see that it has sufficient performance for the production of low integrated DNA chip consisted of 96 spots within $1cm^2$ area.

DNA Marker를 이용한 한국 재래닭의 유전특성 분석 (Analysis of Genetic Characteristics of Korean Native Chicken Using DNA Marker)

  • 이학교;이성진;황규춘;정일정;박용호;손시환;신영수;오봉국;한재용
    • 한국가금학회지
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    • 제23권4호
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    • pp.177-183
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    • 1996
  • This study was conducted to analyze genetic characteristics of Korean Native Chicken three lines classified on the basis of the feather color and appearance (Red, Yellow, and Black) using DNA fingerprinting method. To estimate the genetic relatedness among breeds and similarities within breeds, we collected blood samples from Korean Native Chicken (KNC), Rhode Island Red (RIR), White Leghorn (WL), and Cornish(CN) and obtained genomic DNA from the blood of 10 individuals randomly selected within the breeds and lines. The genomic DNA samples were digested with restriction enzymes (Hinf J, Hae Ill) and hybridized with various probes (Jeffreys' probes 33.15, 33.6 and M13) after Southern transfer. Genetic similarities within breeds were characterized by band sharing (BS) value, estimated by the DFP band pattern between the pair of lanes. BS values within WL, RIR, and KNC were 0.82, 0.70 and 0.56, respectively. Relative genetic diversity (BS value) of KNC was higher than those two breeds (WL, RIR). Estimation of genetic similarity between KNC lines and control breed (RIR) was 0.32, whereas similarity within KNC lines (6 groups) was 0.50. In this analysis, KNC was showed to have a highly genetic diver-sity at the DNA level, and to be closer in genetic distance to RIR (0.67) than any other breeds.

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장환형 단일가닥 DNA를 이용한 암세포 성장 억제 유전자 발굴 (Large-Circular Single-stranded Sense and Antisense DNA for Identification of Cancer-Related Genes)

  • 배윤위;문익재;서영배;도경오
    • 한국미생물·생명공학회지
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    • 제38권1호
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    • pp.70-76
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    • 2010
  • The single-stranded large circular (LC)-sense DNA were utilized as probes for DNA chip experiments. The microarray experiment using LC-sense DNA probes found differentially expressed genes in A549 cells as compared to WI38VA13 cells, and microarray data were well-correlated with data acquired from quantitative real-time RT-PCR. A 5K LC-sense DNA microarray was prepared, and the repeated experiments and dye swap test showed consistent expression patterns. Subsequent functional analysis using LC-antisense library of overexpressed genes identified several genes involved in A549 cell growth. These experiments demonstrated proper feature of LC-sense molecules as probe DNA for microarray and the potential utility of the combination of LC-sense microarray and antisense libraries for an effective functional validation of genes.

Detection of Avian Influenza-DNA Hybridization Using Wavelength-scanning Surface Plasmon Resonance Biosensor

  • Kim, Shin-Ae;Kim, Sung-June;Lee, Sang-Hun;Park, Tai-Hyun;Byun, Kyung-Min;Kim, Sung-Guk;Shuler, Michael L.
    • Journal of the Optical Society of Korea
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    • 제13권3호
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    • pp.392-397
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    • 2009
  • We designed a wavelength interrogation-based surface plasmon resonance (SPR) biosensor to detect avian influenza DNA (AI-DNA). Hybridization reactions between target AI-DNA probes and capture probes immobilized on a gold surface were monitored quantitatively by measuring the resonance wavelength in the visible waveband. The experimental results were consistent with numerical calculations. Although the SPR detection technique does not require the DNA to be labeled, we also evaluated fluorescently-labeled targets to verify the hybridization behavior of the AI-DNA. Changes in resonance were found to be linearly proportional to the amount of bound analyte. A wavelength interrogation-type SPR biosensor can be used for rapid measurement and high-throughput detection of highly pathogenic AI viruses.