• Title/Summary/Keyword: DNA 고정화

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Development of an SH-SAW sensor for detection of DNA (DNA 측정용 SH-SAW 센서 개발)

  • Hur Youngjune;Seon Jooheon;Roh Yongrae
    • Proceedings of the Acoustical Society of Korea Conference
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    • spring
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    • pp.319-322
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    • 2004
  • 본 연구에서는 DNA의 상보적인 결합을 이용하여 DNA 혼성화 반응을 감지할 수 있는 SH형 SAW 센서를 개발하였다. 측정에 사용된 DNA는 15개의 염기를 가진 올리고 뉴클레오티드를 사용하였으며 이에 대해 상보적 결합이 가능한 염기서열을 가진 것과 그렇지 않은 미스매치 형태의 DNA 올리고뉴클레오티드를 이용하여 DNA 혼성화 반응 특성을 측정하였다. SH형 SAW 센서는 압전 단결정 $LiTaO_{3}$를 사용하여 100 MHz 발진되는 형태로 제작하였으며, 센서의 지연선 위에 Ti/Au 층을 증착하여 SH기가 수식된 탐침 DNA의 고정화가 가능하게 하였다. 제작된 센서는 Au가 증착된 박막위에 탐침 DNA를 SAM 방법으로 고정화 시켰을 경우와 고정화된 탐침 DNA와 표적 DNA와의 혼성화 반응을 시키고 난 후의 센서의 주파수 변화를 각각 측정하였다. 개발된 DNA 혼성화 반응 측정용 SH형 SAW센서는 DNA 혼성화 특성에 기인한 질량하중 효과에 따른 안정적인 주파수 변화를 나타내었다.

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Development of an SH-SAW Sensor for Detection of DNA (DNA 측정용 SH-SAW 센서 개발)

  • Hur Youngjune;Pak Yukeun Eugene;Roh Yongrae
    • The Journal of the Acoustical Society of Korea
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    • v.24 no.3
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    • pp.160-165
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    • 2005
  • We have developed SH (shear horizontal) surface acoustic wave (SAW) sensors for detection of the immobilization and hybridization of DNA (deoxyribonucleic acid) on the gold coated delay line of transverse SAW devices. The experiments of DNA immobilization and hybridization were performed with 15-mer oligonucleotides (probe and complementary target DNA). The sensor consists of twin SAW delay line oscillators operating at 100 MHz fabricated on $36^{\circ}$ rotated Y-cut $LiTaO_3$ piezoelectric single crystals. The relative change in the frequency of the two oscillators was monitored to detect the hybridization between target DNA and immobilized probe DNA in pH 7.4 PBS (phosphate buffered saline) solution. The measurement results showed a good response of the sensor to the mass loading effects of the DNA immobilization and hybridization with the sensitivity up to $1.55{\cal}ng/{\cal}ml/Hz$.

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.

Covalent Binding of DNA onto Glass Support for the Construction of Genosensor

  • Jeong, U-Seong;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.709-710
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    • 2000
  • Genosensor technology utilizes a patterned array of DNA molecules immobilized on solid supports for biomedical analysis. The detection capability of the sensor depended mainly on the way the capture probes are attached to the support as well as the sequence. We compared two different. coupling methods currently used to covalently graft DNA molecules onto a glass surface.

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Development of DNA Probe Assay System for Salmonella Species using Glass as substrate

  • Jeong, U-Seong;Lee, Ung-Hui;Baek, Se-Hwan
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.235-236
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    • 2001
  • We developed a DNA probe analytical system with a patterned array of oligonucleotide molecules immobilized on glass surfaces. The detection capability of the system depended mainly on the way the capture probes were attached to the support as wen as the sequence. We optimized major variables to graft DNA molecules onto a glass support and the DNA probe assay was eventually accomplished without purification of the PCR product.

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DNA Selective Immobilization on a Microcantilever with Nano-Interdigitated Electrodes (Nano-IDEs) Using Cyclic Voltammetry (맞물린 나노전극을 가지는 마이크로 캔틸레버의 제작 및 순환전압전류방법을 이용한 DNA의 선택적인 고정화)

  • Lee, Jung-A;Lee, Kwang-Cheol
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.32 no.6
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    • pp.459-464
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    • 2008
  • We present a novel microcantilever device with nano-interdigitated electrodes (nano-IDEs) and DNA selective immobilization on the nano-IDEs for biosensing applications. Using the nano-IDEs and cyclic voltammetric methods, we have achieved selective immobilization of DNA with submicrometer spatial resolution on a freestanding microcantilever. $70{\sim}500\;nm$-wide gold (Au) nano-IDEs are fabricated on a low-stress SiNx microcantilever with dimensions of $100{\sim}600\;{\mu}m$ in length, and $15{\sim}60\;{\mu}m$ in width, with a $0.5\;{\mu}m$ thickness using electron beam lithography and bulk micromachining. Streptavidin is selectively deposited on one side of the nano-IDEs using cyclic voltammetry at a scan rate of 0.1 V/s with a range of $-0.2{\sim}0.7\;V$ during $1{\sim}5$ cycles. The selective deposition of dsDNA is confirmed by fluorescence microscopy after labeling with YOYO-1 dye.

Surface Acoustic Wave Characteristics of Piezoelectric Materials and Protein Immobilization (압전 재료의 탄성표면파 특성과 단백질의 고정화)

  • Chong, Woo-Suk;Hong, Chul-Un;Kim, Gi-Beum
    • Korean Chemical Engineering Research
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    • v.44 no.2
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    • pp.166-171
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    • 2006
  • In this study, in using a piezoelectric material of $Pb(Mg_{1/3}Nb_{2/3})O_3-PbTiO_3$ (PMN-PT), which has a high electromechanical coupling coefficient, we have tried to study about this material can be practically available as a new biosensor to detect protein by using surface acoustic wave (SAW). As the results, the filtering of the center frequency of the PMN-PT substrate is a superior result to that of the $LiTaO_3$ (LT) substrate, but the result was not completely satisfactory. Also this study attempts to develop a sensing method to detect mismatched DNA in order to diagnose cancer. We could directly immobilize the MutS to the NTA using the EDC solution. But, we immobilized MutS using nickel and it is judged that is more effective method to detect mismatched DNA.

A Study on the Immobilization of Biomolecules on Poly(acrylic acid)-grafted MWCNTs Prepared by Radiation-Induced Graft Polymerization (방사선 그래프트 중합에 의하여 제조된 폴리(아크릴 산)이 그래프트된 탄소나노튜브에 생체분자 고정화에 관한 연구)

  • Jung, Chan-Hee;Lee, Byoung-Min;Hwang, In-Tae;Choi, Jae-Hak;Nho, Young-Chang;Hong, Sung-Kwon
    • Polymer(Korea)
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    • v.34 no.2
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    • pp.150-153
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    • 2010
  • In this research, biomolecule-immobilized multi-walled carbon nanotubes (MWCNTs) were prepared by using radiation-induced graft polymerization. For the immobilization of biomolecules, the surface of MWNCTs was functionalized by radiation-induced graft polymerization of acrylic acid. Based on the results of TGA and Raman spectroscopy it was found that acrylic acid was effectively graft-polymerized on the MWCNTs. Biomolecules such as DNA and proteins were immobilized onto the resultant poly(acrylic acid)-grafted MWCNTs. The results of the X-ray photoelectron spectroscopy and fluorescence microscopy confirmed that the biomoelcules were successfully immobilized on the poly(acrylic acid)-grafted MWCNTs.

Growth Promotion of Tomato by Application of Immobilized Arthrobacter woluwensis ED in Alginate Beads (Alginate에 고정화된 Arthrobacter woluwensis ED 처리 시 토마토의 생장촉진과 균주의 토양 내 잔류)

  • Kwon, Seung-Tak;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.40-45
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    • 2014
  • In order to increase the persistence of plant growth promoting rhizobacteria (PGPR) in rhizpsphere soil, the growth of tomato was examined after the application of Arthrobacter woluwensis ED immobilized in alginate bead, which was known as PGPR. When tomato seedlings were treated with A. woluwensis ED of $1{\times}10^6$ cells g $soil^{-1}$ and incubated for 30 days in a plant growth chamber, the shoot length, root length, fresh weight and dry weight of the grown tomato plants treated with the suspended inoculants significantly increased by 36.2, 59, 51.1, and 37.5%, respectively compared to those of the uninoculated control. The treatment of the immobilized bacteria increased those by 42, 67.4, 62.5, and 60.4%, respectively compared to those of the uninoculated control. Therefore, the enhancement of tomato growth by the treatment of the immobilized bacteria was higher than those by the suspended inoculants. The effects of the inoculation on indigenous bacterial community and the fate of the inoculated bacteria were monitored by denaturing gradient gel electrophoresis analysis. The DNA band intensity of A. woluwensis ED in the tomato rhizosphere treated with the suspended inoculants continuously decreased after the inoculation, but the band intensity in the tomato rhizosphere soils treated with the immobilized inoculants showed the maximum at 1 week after inoculation and the decreasing rate was less than that of the suspended inoculants, which indicated the longer maintenance of the immobilized bacteria at rhizosphere soils. Therefore, encapsulation of PGPR in alginate beads may be more effective than liquid inoculant for the plant growth promotion and survival of PGPR at plant rhizosphere.

Effect of PVA-Encapsulation on Hydrogen Production and Bacterial Community Structure (수소 생산과 세균 군집구조에 미치는 PVA-포괄고정화의 영향)

  • Yun, Jeonghee;Kim, Tae Gwan;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.41-50
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    • 2014
  • In this study, the performances of PVA-encapsulation and non-encapsulation in a fed-batch bioreactor system were compared for biohydrogen production. Hydrogen production in the PVA-encapsulation bioreactor was not significantly different in comparison to the non-encapsulation bioreactor. However, the hydrogen gas in the encapsulation bioreactor could be stably produced when it was exposed to environmental difficulties such as pH impact by the accumulation of organic acids as fermentative metabolic products. Bacterial communities by DGGE analysis were differently shifted between the PVA-encapsulation and non-encapsulation bioreactors from the initial sludge. The community of hydrogen producing bacteria was stable during the experimental period in the PVA-encapsulation bioreactor compared to the non-encapsulation method. The absolute quantitation of the DNA copy number by a high-throughput droplet digital PCR system for six genera contributed to hydrogen production showing that the numbers of dominant bacteria existed at similar levels in the two bioreactors regardless of encapsulation. In both of two bioreactors, not only Clostridium and Enterobacter, which are known as anaerobic hydrogen producing bacteria, but also Firmicutes, Ruminococcus and Escherichia existed with $1{\times}10^5-1{\times}10^6$ copy numbers of ml-samples exhibiting rapid growth during the initial operation period.