• Title/Summary/Keyword: microchannel electrophoresis

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Recent Advances in DNA Sequencing by End-labeled Free-Solution Electrophoresis (ELFSE)

  • Won, Jong-In
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.179-186
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    • 2006
  • End-Labeled Free-Solution Electrophoresis (ELFSE) is a new technique that is a promising bioconjugate method for DNA sequencing (or separation) and genotyping by both capillary and microfluidic device electrophoresis. Because ELFSE enables high-resolution electrophoretic separation in aqueous buffer alone (i.e., without a polymer matrix), it eliminates the need to load viscous polymer networks into electrophoresis microchannels. To achieve microchannel DNA separations with high performance, ELFSE requires monodisperse perturbing entities (i.e., drag-tags), which create a large amount of frictional drag when pulled behind DNA during free-solution electrophoresis, and which have other properties suitable for microchannel electrophoresis. In this article, the theoretical concepts of ELFSE and the required characteristics of the drag-tag molecules for the ultimate performance of ELFSE are reviewed. Additionally, the merits and limitations of current drag-tags are also discussed in the context of recent experimental data of ELFSE separation (or sequencing).

PDMS/Glass Based DNA Microbiochip for Restriction Enzyme Reaction and Electrophoresis Detection (DNA의 제한효소 반응 및 전기영동 검출용 PDMS/유리 마이크로바이오칩)

  • Choi Joon-Young;Ahn Yoo Min;Hwang Seung-Yong
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.30 no.1 s.244
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    • pp.26-31
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    • 2006
  • This paper reports low-cost PDMS/glass based DNA microbiochip for the restriction enzyme reaction and its products detection using the capillary electrophoresis. The microbiochip ($25mm{\times}75mm$) has the heater integrated reactor ($5{\mu}{\ell}$) for DNA restriction enzyme reaction at $37^{\circ}C$ and the microchannel ($80\;{\mu}m{\times}100\;{\mu}m{\times}58mm$) for the capillary electrophoresis detection. It is experimentally confirmed that the digestion of the plasmid ($pGEM^{(R)}-4Z$) by the enzyme (Hind III and Sca I) is performed for less than 10 min and its electrophoresis detection is able to sequentially on the fabricated microbiochip.

Evaluation of Electrokinetic Flow Mobility Using Isotacho-Electrophoresis Techniques

  • An, J.H.;Joo, Y.H.;Lee, C.Y.;Lee, Y.J.;Park, C.W.
    • Journal of Magnetics
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    • v.16 no.4
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    • pp.444-448
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    • 2011
  • In the present study, we separated the marker particles from the suspending particle mixture solution using isotacho-electrophoresis technique, a novel quantitative ionic particle separation method, in the microchannel. A multiple stacking zone of the suspending particle was visualized with variations in electric field strength, pH value and concentration of the ionic solution. In particular, the electrophoretic mobility of ionic particle (fluorescein) was estimated based on the electrophoretic velocity value measured by the particle image velocimetry. As a result, isotacho-electrophoresis zones were clearly visualized as going downstream in the electric field. The particle migration velocity increased proportional to the applied voltage increase; it was also affected by the pH value variations in the ionic solution.

Dielectric Interaction of Particle in Electrophoresis (전기 영동에서의 입자간의 전기력에 의한 상호 작용)

  • Lee, Ho-Rim;Kang, Kwan-Hyoung
    • 유체기계공업학회:학술대회논문집
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    • 2006.08a
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    • pp.171-174
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    • 2006
  • When two particles close to each other are in electrophoretic motion, each particle is under the influence of the non-uniform electric field generated by the other particle. Two particles may attract or repel each other due to the dielectric force depending on their positions in the non-uniform electric field. It is shown analytically that two adjusting rigid particles can form an aggregate due to the dielectric interaction. To verify the validity of the theoretical prediction, an experiment is carried out by using a microchannel. In the experiment, AC electric field is used to eliminate cumbersome electroosmotic flow. The experimental result shows that the particles form a chain-like structure, which is typically observed in electro-rheological fluid, due to the dielectric interaction.

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MEMS Fabrication of Microchannel with Poly-Si Layer for Application to Microchip Electrophoresis (마이크로 칩 전기영동에 응용하기 위한 다결정 실리콘 층이 형성된 마이크로 채널의 MEMS 가공 제작)

  • Kim, Tae-Ha;Kim, Da-Young;Chun, Myung-Suk;Lee, Sang-Soon
    • Korean Chemical Engineering Research
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    • v.44 no.5
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    • pp.513-519
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    • 2006
  • We developed two kinds of the microchip for application to electrophoresis based on both glass and quartz employing the MEMS fabrications. The poly-Si layer deposited onto the bonding interface apart from channel regions can play a role as the optical slit cutting off the stray light in order to concentrate the UV ray, from which it is possible to improve the signal-to-noise (S/N) ratio of the detection on a chip. In the glass chip, the deposited poly-Si layer had an important function of the etch mask and provided the bonding surface properly enabling the anodic bonding. The glass wafer including more impurities than quartz one results in the higher surface roughness of the channel wall, which affects subsequently on the microflow behavior of the sample solutions. In order to solve this problem, we prepared here the mixed etchant consisting HF and $NH_4F$ solutions, by which the surface roughness was reduced. Both the shape and the dimension of each channel were observed, and the electroosmotic flow velocities were measured as 0.5 mm/s for quartz and 0.36 mm/s for glass channel by implementing the microchip electrophoresis. Applying the optical slit with poly-Si layer provides that the S/N ratio of the peak is increased as ca. 2 times for quartz chip and ca. 3 times for glass chip. The maximum UV absorbance is also enhanced with ca. 1.6 and 1.7 times, respectively.

Fabrication of electro phoresis microchips and effects of channel surface properties (마이크로 전기영동 소자의 제작과 유로 면 특성에 따른 전기삼투 및 전기영동 효과)

  • Kim, Min-Su;Cho, Seung-Il;Lee, Kook-Nyung;Kim, Yong-Kweon
    • Proceedings of the KIEE Conference
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    • 2003.10a
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    • pp.286-289
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    • 2003
  • We investigated the influence of the properties of substrate material on the separation efficiency in microchip electrophoresis. We fabricated the various microchips and studied separation efficiency in microchannels composed of a single material such as quartz, glass, polydimethylsiloxane (PDMS), and polymethylmetha crylate (PMMA), as well as hybrid micro channels composed of different materials. New fabrication process for glass chip was suggested and some treatment is added to improve fabrication process in other chip. Separation efficiency was compared by measuring migration times and bandwidths of EOF and analytes in each microchip. The efficiency is the function of migration time, which is affected by the electroosmotic flow (EOF), and bandwidth of an analyte. EOF is highly dependent upon the characteristics of a microchannel wall surface. Migration time was more reproducible in silica chips than that of PDMS chip and more band broadening was observed in the microchip composed of hybrid material due to non-uniformity of surface charge density at the walls of the channel.

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An integrated DNA barcode assay microdevice for rapid, highly sensitive and multiplex pathogen detection at the single-cell level

  • Jung, Jae Hwan;Cho, Min Kyung;Chung, So Yi;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.276-276
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    • 2013
  • Here we report an integrated microdevice consisting of an efficient passive mixer, a magnetic separation chamber, and a capillary electrophoretic microchannel in which DNA barcode assay, target pathogen separation, and barcode DNA capillary electrophoretic analysis were performed sequentially within 30 min for multiplex pathogen detection at the single-cell level. The intestine-shaped serpentine 3D micromixer provides a high mixing rate to generate magnetic particle-pathogenic bacteria-DNA barcode labelled AuNP complexes quantitatively. After magnetic separation and purification of those complexes, the barcode DNA strands were released and analyzed by the microfluidic capillary electrophoresis within 5 min. The size of the barcode DNA strand was controlled depending on the target bacteria (Staphylococcus aureus, Escherichia coli O157:H7, and Salmonella typhimurium), and the different elution time of the barcode DNA peak in the electropherogram allows us to recognize the target pathogen with ease in the monoplex as well as in the multiplex analysis. In addition, the quantity of the DNA barcode strand (~104) per AuNP is enough to be observed in the laser-induced confocal fluorescence detector, thereby making single-cell analysis possible. This novel integrated microdevice enables us to perform rapid, sensitive, and multiplex pathogen detection with sample-in-answer-out capability to be applied for biosafety testing, environmental screening, and clinical trials.

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