• Title/Summary/Keyword: DNA 분리 칩

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Amperometric detection of DNA using capillary electrophoresis on microchip (모세관 전기영동 마이크로칩을 이용한 디옥시리보핵산(DNA)의 전류법 검출)

  • Joo, Gi-Sung;Ha, Kon;Jha, Sandeep K.;Lee, Hyun-Ho;Yoon, Tae-Sik;Kang, C.J.;Kim, Yong-Sang
    • Proceedings of the KIEE Conference
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    • 2008.07a
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    • pp.1460-1461
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    • 2008
  • 마이크로칩 형태에서의 모세관 전기영동과 전류법을 이용하여 디옥시리보핵산(DNA) 단편들의 분리 검출하는 실험을 하였다. 마이크로 채널이 형성된 PDMS(polydimethylsiloxane)와 금 전극이 형성된 유리 기판을 접합하여 마이크로칩을 제작하였다. 20V/cm의 전계를 인가하여 100bp-1.5kbp 길이의 DNA 단편을 모세관 전기영동 하였을 때 250초내에 분리 검출되는 것을 확인하였다.

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Development of Exposure Biomarkers for Endocrine Disrupting Chemicals Using DNA Microarray (DNA 마이크로어레이를 이용한 내분비장애물질 노출지표 개발)

  • Yang, Mi-Hi
    • Environmental Analysis Health and Toxicology
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    • v.20 no.4 s.51
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    • pp.327-332
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    • 2005
  • 장기간 노출 시 발암 등 인체 유해성을 갖는 환경유래 내분비장애물질(endocrine disrupting chemicals, EDCs)에 대한 선택적이고 민감한 노출지표를 개발하기 위하여 본 연구에서는 DNA microarray를 이용하였다. 피험자는 아직 특별한 질환을 갖지 않는 18세 이상 연령, 성을 맞춘 EDCs고농도 노출군(N = 16)과 저농도군(N = 16)으로 구성되었다. 노출정도 구분은 10년 이상 거주지가 K산업폐기물 소각장과 2.5 km 반경 내, 외 인지에 따라 고노출군,저노출군으로 구분하였다. 피험자의 말초혈에서 total RNA를 분리, 각 군당 B인씩 pool로 cDNA를 합성하여 oligonucleotide DNA 칩에 적용하였다. 유전자발현의 차이를 GenePixPro 4.0 software를 이용하여 분석하였다. 총 3장의 칩을 이용하여 공통적으로 저노출군보다 고노출군에서 2배 이상 발현의 증가를 보인 유전자는 plasminogen activator(PLAT)등 12종이 관찰되었고, l/2이하로 발현의 감소를 보인 유전자는 kallikrein 3 (KLK3)등 29종이었다. 이 들 유전자는 PLAT등 면역계 반응에 관여하는 유전자 및 apoptosis, transport, G protein, chromatin, 암화, 발생 (development), 대사 등에 관여하는 유전자들이었다. 그러므로 KLK3등 본 연구에서 발굴한 유전자는 향후 확대된 인구에서 본 연구 결과의 확인을 통하여 EDCs특이적 노출지표로써, 나아가 암 등 EDCs관련 질병의 기전 및 병인학을 구명하는데 이용가치가 높다고 사료된다.

Development of the DNA Sequencing Chip with Nano Pillar Array using Injection Molding (Nano Pillar Array 사출성형을 이용한 DNA 분리 칩 개발)

  • Kim S.K.;Choi D.S.;Yoo Y.E.;Je T.J.;Kim T.H.;Whang K.Y.
    • Proceedings of the Korean Society of Precision Engineering Conference
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    • 2005.06a
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    • pp.1206-1209
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    • 2005
  • In recent, injection molding process for features in sub-micron scale is under active development as patterning nano-scale features, which can provide the master or stamp for molding, and becomes available around the world. Injection molding has been one of the most efficient processes for mass production of the plastic product, and this process is already applied to nano-technology products successfully such as optical storage media like DVD or BD which is a large area plastic thin substrate with nano-scale features on its surface. Bio chip for like DNA sequencing may be another application of this plastic substrate. The DNA can be sequenced using order of 100 nm pore structure when making the DNA flow through the pore structure. Agarose gel and silicon based chip have been used to sequence the DNA, but injection molded plastic chip may have benefit in terms of cost. This plastic DNA sequencing chip has plenty of pillars in order of 100 nm in diameter on the substrate. When the usual features in case of DVD or BD have very low aspect ratio, even less than 0.5, but the DNA chip will have relatively high aspect ratio of about 2. It is not easy to injection mold the large area thin substrate with sub-micron features on its surface due to the characteristics of the molding process and it becomes much more difficult when the aspect ratio of the features becomes high. We investigated the effect of the molding parameters for injection molding with high aspect ratio nano-scale features and injection molded some plastic DNA sequencing chips. We also fabricated PR masters and Ni stamps of the DNA chip to be used for molding

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High-Speed RNA Isolation Using Magnetic Oligo(dT) Beads and Lateral Magnetophoresis (올리고-dT 자성입자와 측면방향 자기영동을 이용한 초고속 RNA 추출 기술)

  • Lee, Hwan-Yong;Han, Song-I;Han, Ki-Ho
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.35 no.12
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    • pp.1309-1316
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    • 2011
  • This paper presents a high-speed RNA microextractor for the direct isolation of RNA from blood lysate using magnetic oligo(dT) beads. The extraction is performed through lateral magnetophoresis, which is induced by a ferromagnetic wire array inlaid. With this RNA microextractor, more than 80% of the magnetic beads could be separated at a flow rate up to 20 ml/h, and the overall extraction procedure was completed within 1 min. The absorbance ratio of RNA to protein(A260/A280) was greater than 1.7, indicating that the extraction technique yields pure RNA. The feasibility of using this technique in reverse transcription polymerase chain reaction procedures was investigated by cDNA synthesis and PCR processes. The results confirmed that the RNA microextractor is a practical device for easy, fast, and high-precision RT-PCR using minimal amounts of reagent.

Detection of Point Mutations in the rpoB Gene Related to Drug Susceptibility in Mycobacterium Tuberculosis using an Oligonucleotide Chip (올리고뉴클레오티드 칩(Oligonucleotide Chip)을 이용한 항결핵제 감수성과 관련된 Mycobacterium tuberculosis rpoB 유전자의 점돌연변이 판별 방법)

  • Kim, Hyun-Jung;Kim, Seong-Keun;Shim, Tae-Sun;Park, Yong-Doo;Park, Mi-Sun
    • Tuberculosis and Respiratory Diseases
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    • v.50 no.1
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    • pp.29-41
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    • 2001
  • Background : The appearance of multiple-drug-resistant Mycobacterium tuberculosis strains has been seriously compromising successful control of tuberculosis. Rifampin-resistance, caused by mutations in the rpoB gene, can be indicative of multiple-drug-resistance, and its detection is of great importance. The present study aimed to develop an oligonucleotide chip for accurate and convenient screening of drug-resistance. Methods : In order to detect point mutations in the rpoB gene, an oligonucleotide chip was prepared by immobilizing specific probe DNA to a microscopic slide glass by a chemical reaction. The probe DNA that was selected from the 81 bp core region of the rpoB gene was designed to have mutation sites at the center. A total of 17 mutant probes related to rifampin-resistance including 8 rifabutin-sensitive mutant probes were used in this study. For accurate determination, wild type probes were prepared for each mutation position with an equal length, which enabled a direct comparison of the hybridization intensities between the mutant and wild type. Results : Mycobacterial genomic DNA from clinical samples was tested with the oligonucleotide chip and the results were compared with those of the drug-susceptibility test in addition to sequencing and INNO-LiPA Rif. TB kit test in some cases. Out of 15 samples, the oligonucleotide chip results of 13 samples showed good agreement with the rifabutin-sensitivity results. The two samples with conflicting result also showed a discrepancy between the other tests, suggesting such possibilities as existence of mixed strains and difference in drug-sensitivity. Further verification of these samples in addition to more case studies are required before the final evaluation of the oligonucleotide chip can be made. Conlcusion : An oligonucleotide chip was developed for the detection of rpoB gene mutations related to drugsusceptibility. The results to date show the potential for using the oligonucleotide chip for accurate and convenient screening of drug-resistance to provide useful information in antituberculosis drug therapy.

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Microarray Analysis of Gene Expression in Rat Hippocampus of Maternal Social Separation Model (모성 및 사회성 분리 백서 모델의 해마에서 유전자 칩을 이용한 유전자 발현 연구)

  • Lee, Hee Jae;Son, Chang Hee;Kwak, Hyong Ryol;Lee, Sang-Hyun;Han, Yoon Hee;Kim, Soo Young;Park, Jong-Ik;Chun, Wanjoo;Kim, Sung-Soo
    • Korean Journal of Biological Psychiatry
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    • v.13 no.2
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    • pp.110-116
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    • 2006
  • Objectives : Alteration of hippocampus was demonstrated in the maternal social separation(MSS) pups, separated from dams on postnatal day(pnd) 14 and placed alone. Therefore, to understand the molecular events involved in the MSS, we have initiated a search for gene profiles that are up or down-regulated in the hippocampus of MSS pups. Methods : Analysis of cDNA microarray was performed by using total RNA extracted from the hippocampus of control and MSS pups on pnd 17. Also, passive-avoidance test was demonstrated on pnd 35. Results : Up-regulation of Nedd4a was observed in the hippocampus of MSS pups. Also, MSS rats showed less elongation of latency in passive avoidance test. Conclusion : We suggest that environmental effects of MSS may be altered the neural and/or glial differentiation and synapse formation-related genes which may lead cognitive alterations in MSS rats.

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Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato (유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색)

  • Shin, Weon-Sun;Kim, Myung-Hee
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.591-597
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    • 2003
  • To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

Diagnosis of the ORF Virus Using a Mixture of Sieving Gel Matrixes in Microchip Gel Electrophoresis (마이크로칩젤 전기영동에서 충진젤 혼합물을 이용한 ORF 바이러스의 진단)

  • Kim, Yun-Jeong;Chae, Joon-Seok;Kang, Seong-Ho
    • Journal of the Korean Chemical Society
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    • v.48 no.5
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    • pp.483-490
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    • 2004
  • We have developed a novel polymerase chain reaction (PCR)-microchip gel electrophoresis (MGE) method based on the sieving gel mixture of commercially available poly(vinylpyrrolidone) (PVP) and hydroxy ethyl cellulose (HEC) for the rapid detection and diagnosis of the orf virus (ORFV) from Korean indigenous goat. After amplification of 594-bp DNA fragment from the B2L gene of ORF virus, the amplicon was analyzed by the MGE separation. The glass microfluidic chip (64 mm total length (36 mm effective length)${\times}$90 ${\mu}$m width${\times}$20 ${\mu}$m depth) allowed the fast detection and diagnosis of ORFV in the mixture of 1.0% PVP ($M_r$ 360,000) and 1.0% HEC ($M_r$250,000) as a sieving matrix with better resolution and reproducibility of DNA fragments. Under the electric field of 277.8 V/cm, the 594-bp DNA was analyzed within 4 min. Compared to traditional slab gel electrophoresis, the PCR-MGE method was twenty times faster and an effective clinical method for the quantitative analysis of ORFV.

Novel Pathogenic Strain of Watermelon mosaic virus Occurred on Insam (Panax ginseng) (인삼(Panax ginseng)에 발생한 Watermelon mosaic virus의 새로운 병원성 계통)

  • Jung, Won-Kwon;Nam, Moon;Lee, Joo Hee;Park, Chung Youl;Kim, Byoung Hoon;Park, Eun Hye;Lee, Min-A;Kim, Mi-Kyeong;Choi, Hong-Soo;Lee, Jun Seong;Kim, Jeong-Soo;Choi, Jin Kook;Kwon, Tae Ryong;Lee, Key-Woon;Lee, Su-Heon
    • Research in Plant Disease
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    • v.19 no.4
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    • pp.331-337
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    • 2013
  • A disease, supposedly caused by a virus, was observed from Insam (Panax ginseng) fields of Punggi in year 2006. It has long believed to be a physiological disorder. However, the incidence of the disease has increased every year. When several samples were observed under electron microscope, filamentous virus-like particles were observed. The nucleotide sequences of the virus were analyzed by RT-PCR with specific primer sets derived from the results of DNA chip. The results indicated that the disease was caused by Watermelon mosaic virus (WMV). It revealed that the result of the biological assay by the virus was different from that of WMV previously found in other crops. Therefore, this is the first report that WMV causes the disease in P. ginseng and the virus is named to be WMV-Insam.

A Report of Five Unrecorded Fungal Species of Korea (국내 미기록 진균 5종 보고)

  • Ahn, Geum Ran;Kim, Bo Young;Lee, Geun Sick;Hyun, Min Woo;Lee, Chan Jung;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.240-246
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    • 2016
  • During a survey of the activities of fungi in steamed sweet potato, stored garlic, agricultural by-products for mushroom cultivation media, and pinewood chips from a pinewood nematode-infected tree, numerous fungal samples were isolated and identified. This study identified five species that have not been previously reported in Korea, namely Geomyces pannorum, Neopestalotiopsis javaensis, Penicillium allii, Penicillium chermesinum, and Ophiognomonia setacea. For all identified species, the cultural features of colonies formed on growth media, their morphological characteristics observed by a light microscope, and their molecular phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer rDNA region or calmodulin gene were described.