• 제목/요약/키워드: DNA Microarray

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DNA Microarray Probe Preparation by Gel Isolation Nested PCR

  • Wang, Hong-Min;Ma, Wen-li;Huang, Hai;Xiao, Wei-Wei;Wang, Yan;Zheng, Wen-Ling
    • BMB Reports
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    • 제37권3호
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    • pp.356-361
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    • 2004
  • To develop a simplified method that can rapidly prepare DNA microarray probes in a massive scale, a lambda phage genomic DNA-fragments library was constructed for the microarray-probes collection. Four methods of DNA band recovery from the first PCR products were tested and compared. The DNA microarray probes were collected by a novel method of nested PCR that was mediated by gel isolation of the first PCR products. This method was named GIN-PCR. The probes that were prepared by this GIN-PCR technique were used as subjects to fabricate a DNA microarray. The results showed that a wooden toothpick was superior to the other 3 methods, since this technique can steadily transfer the DNA bands as the template of the second PCR after the first PCR. A group of probes were successfully collected and DNA microarrays were constructed using these probes. Hybridization results demonstrated that this technique of DNA recovery and probe preparation was rapid, efficient, and effective. We developed a cost-effective and less labor-intensive method for DNA microarray probe preparation by nested PCR that is mediated by wooden toothpick transfer of the DNA bands in the gel after electrophoresis.

Evaluation of Amplified-based Target Preparation Strategies for Toxicogenomics Study : cDNA versus cRNA

  • Nam, Suk-Woo;Lee, Jung-Young
    • Molecular & Cellular Toxicology
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    • 제1권2호
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    • pp.92-98
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    • 2005
  • DNA microarray analysis of gene expression in toxicogenomics typically requires relatively large amounts of total RNA. This limits the use of DNA microarray when the sample available is small. To confront this limitation, different methods of linear RNA amplification that generate antisense RNA (aRNA) have been optimized for microarray use. The target preparation strategy using amplified RNA in DNA microarray protocol can be divided into direct-incorporation labeling which resulted in cDNA targets (Cy-dye labeled cDNA from aRNA) and indirect-labeling which resulted in cRNA targets (i.e. Cy-dye labeled aRNA), respectively. However, despite the common use of amplified targets (cDNA or cRNA) from aRNAs, no systemic assessment for the use of amplified targets and bias in terms of hybridization performance has been reported. In this investigation, we have compared the hybridization performance of cRNA targets with cDNA targets from aRNA on a 10 K cDNA microarrays. Under optimized hybridization conditions, we found that 43% of outliers from cDNA technique and 86% from the outlier genes were reproducibly detected by both targets hybridization onto cDNA microarray. This suggests that the cRNA labeling method may have a reduced capacity for detecting the differential gene expression when compared to the cDNA target preparation. However, further validation of this discordant result should be pursued to determine which techniques possesses better accuracy in identifying truly differential genes.

DNA Chip Microarrays를 위한 template로서 소수성 패턴의 제작 (Fabrication of Hydrophobic/Hydrophilic Pattern as a Template for DNA Chip Microaray)

  • 최용성;박대희
    • 한국전기전자재료학회:학술대회논문집
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    • 한국전기전자재료학회 2004년도 추계학술대회 논문집 Vol.17
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    • pp.472-475
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    • 2004
  • Microarray-based DNA chips provide an architecture for multi-analyte sensing. In this paper, we report a new approach for DNA chip microarray fabrication. Multifunctional DNA chip microarray was made by immobilizing many kinds of biomaterials on transducers (particles). DNA chip microarray was prepared by randomly distributing a mixture of the particles on a chip pattern containing thousands of m-scale sites. The particles occupied a different sites from site to site. The particles were arranged on the chip pattern by the random fluidic self-assembly (RFSA) method, using a hydrophobic interaction for assembly.

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유전 알고리즘을 이용한 DNA Microarray의 Probe 선택 (Probe Selection of DNA Microarrays Using Genetic Algorithms)

  • 김선;장병탁
    • 한국지능시스템학회:학술대회논문집
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    • 한국퍼지및지능시스템학회 2002년도 춘계학술대회 및 임시총회
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    • pp.183-187
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    • 2002
  • DNA microarray는 분자생물학 및 DNA 컴퓨팅 분야에 널리 사용되고 있는 실험 도구이다. DNA microarray를 이용하는 한 예는 알려진 유전자 집합을 바탕으로 하여 hybridization을 통해 새로운 DNA 서열을 분석하는 것이다. 이를 위한 가장 간단한 방법은 알려진 유전자의 모든 서열을 DNA microarray 상에 올려놓는 것이지만 이는 결과의 정확도 및 칩 제작비용 면에서 비효율적이다. 따라서 일반적으로는 유전자 서열 정보를 파악한 후 일련의 DNA 서열을 선택하는 probe 디자인 과정을 거친다. 그러나 현재 유전자 서열을 바탕으로 최적의 probe 집합을 찾는 결정적인 방법이 존재하고 있지 않다. 이에 본 논문은 oligo DNA microarray을 이용한 DNA 서열 분석 문제에 있어서 가능한 많은 유전자를 인식하면서 최소의 probe 개수를 갖는 집합을 찾는 방법을 제안한다. 제시된 방법은 가능한 probe 집합들로 해집합을 구성한 후, 유전알고리즘을 이용한 진화 과정을 통해 목적하는 probe 집합을 찾는다. 본 논문에서는 GenBank로부터 얻은 일련의 유전자 집합을 대상으로 실험하였으며 그 결과를 분석하였다.

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Network-based Microarray Data Analysis Tool

  • Park, Hee-Chang;Ryu, Ki-Hyun
    • Journal of the Korean Data and Information Science Society
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    • 제17권1호
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    • pp.53-62
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    • 2006
  • DNA microarray data analysis is a new technology to investigate the expression levels of thousands of genes simultaneously. Since DNA microarray data structures are various and complicative, the data are generally stored in databases for approaching to and controlling the data effectively. But we have some difficulties to analyze and control the data when the data are stored in the several database management systems or that the data are stored to the file format. The existing analysis tools for DNA microarray data have many difficult problems by complicated instructions, and dependency on data types and operating system. In this paper, we design and implement network-based analysis tool for obtaining to useful information from DNA microarray data. When we use this tool, we can analyze effectively DNA microarray data without special knowledge and education for data types and analytical methods.

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소수성 Template를 이용한 DNA Chip Microarray의 개발 (Development of DNA Chip Microarray Using Hydrophobic Template)

  • 최용성;박대희
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2004년도 추계학술대회 논문집 전기물성,응용부문
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    • pp.271-274
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    • 2004
  • Microarray-based DNA chips provide an architecture for multi-analyte sensing. In this paper, we report a new approach for DNA chip microarray fabrication. Multifunctional DNA chip microarray was made by immobilizing many kinds of biomaterials on transducers (particles). DNA chip microarray was prepared by randomly distributing a mixture of the particles on a chip pattern containing thousands of m-scale sites. The particles occupied a different sites from site to site. The particles were arranged on the chip pattern by the random fluidic self-assembly (RFSA) method, using a hydrophobic interaction for assembly.

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DNA microarray를 이용한 항진균 활성세균 Bacillus lentimorbus WJ5의 유전자 발현 분석 (DNA Microarray Analysis of Gene Expression in Antifungal Bacterium of Bacillus lentimorbus WJ5)

  • 이영근;김재성;장유신;조규성;장화형
    • 미생물학회지
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    • 제39권3호
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    • pp.141-147
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    • 2003
  • 여러 항진균 활성 관련 유전자들의 발현 수준을 동시에 연구하기 위하여 DNA microarray를 이용하여 유전자들의 발현 패턴을 비교 분석하였다. 본 연구에서는 항진균활성을 가지는Bacillus lentimorbus WJ5의 genomic DNA를 무작위 하게 제한효소로 절단하여 2,000개의 DNA단편을 microarray하였으며, 감마선($^{60}Co$)조사로 유도된 7종의 항진균 활성 결핍 돌연변이체와 발현양상을 정량적으로 비교하였다. Gene Cluster (Michael Risen, Stanford Uniy.)를 이용한 DNA microarray의 분석 결과, 총 408개의 DNA 단편이 발현되는 것을 확인할 수 있었으며, 이들 중 20개의 DNA단편이 항진균 활성 결핍 돌연변이체에서 발현이 억제되는 것으로 나타났다. 특히,pbuX (xanthine permease, K222), ywbA (phosphotransferase system enzyme II, K393), ptsG (PTS glucose specific enzyme II ABC component, K877), yufO (ABC transporter(ATP-binding protein), K1301), 그리고 ftsY (signal recognition particle (docking protein), K868)는 모든 돌연변이체에서 동시에 발현되는 down-regulation된 유전자들로서 물질 이동과 관련된 것으로 보고되어 있으며, 항진균 활성 관련 신호 및 물질의 이동에 관여할 것으로 사료되어진다.

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.

DNA Microarray 발현정보에 대한 생물학적 정보처리에 관한 연구 (A Study of a Biological Information Processing for DNA Microarray Expression Data)

  • 조영임;정현철
    • 한국지능시스템학회:학술대회논문집
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    • 한국지능시스템학회 2007년도 추계학술대회 학술발표 논문집
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    • pp.149-152
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    • 2007
  • 본 논문은 바이오 인포메틱스의 분야를 간단히 소개하고 기능유전체학에서 microarray 실험에 대한 통계적 방법론을 살펴보고자 한다. 또한 DNA chip 설계와 생물학적 특정에 대해 살펴보고 각 분야에서 적용되는 통계적 방법을 연구분석 해보고자 한다.

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Genes expression monitoring using cDNA microarray: Protocol and Application

  • Muramatsu Masa-aki
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2000년도 국제심포지움 및 추계학술대회
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    • pp.31-41
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    • 2000
  • The major issue in the post genome sequencing era is determination of gene expression patterns in variety of biological systems. A microarray system is a powerful technology for analyzing the expression profile of thousands of genes at one experiment. In this study, we constructed cDNA microarray which carries 2,304 cDNAS derived from oligo-capped mouse cDNA library. Using this hand-made microarray we determined gene expression in various biological systems. To determine tissue specific genes, we compared Nine genes were highly-expressed in adult mouse brain compared to kidney, liver, and skeletal muscle. Tissue distribution analysis using DNA microarray extracted 9 genes that were predominantly expressed in the brain. A database search showed that five of the 9 genes, MBP, SC1, HiAT3, S100 protein-beta, and SNAP25, were previously known to be expressed at high level in the brain and in the nervous system. One gene was highly sequence similar to rat S-Rex-s/human NSP-C, suggesting that the gene is a mouse homologue. The remaining three genes did not match to known genes in the GenBank/EMBL database, indicating that these are novel genes highly-expressed in the brain. Our DNA microarray was also used to detect differentiation specific genes, hormone dependent genes, and transcription-factor-induced genes. We conclude that DNA microarray is an excellent tool for identifying differentially expressed genes.

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