• 제목/요약/키워드: immobilized oligonucleotide probe

검색결과 8건 처리시간 0.032초

Determination of Tyrosinase mRNA in Melanoma by Reverse Transcription-PCR and Optical Mirror Resonance Biosensor

  • Taeboo Choe;Park, Inchul;Seokil Hong
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제7권4호
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    • pp.212-215
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    • 2002
  • Tyrosinase transcript In the blood Is known as the marker of malignant melanoma and it has been often determined by using reverse transcription-polymerase chain reaction (RT-PCA) . However, after the PCR process, the quantification of amplified CDMA by the gel electrophoresis is not reliable and time-consuming. for this reason, we tried to quantify the PCR product using a cuvette-type biosensor, where the oligonucleotide probe was immobilized on the cuvette surface and the single strand CDMA, the denatured PCH product, was then hybridized onto the immobilized probe to give a response signal. The response was Immediate and takes 15 min to obtain a stable signal. The biosensor was much more sensitive comparing to the gel electrophoresis method. The quantification of PCR product using a cuvette-type biosensor was feasible and rapid.

DNA Hybridization 검출 센서를 이용한 매치 및 미스매치 DNA hybridization 특성 연구 (A Study on Match and Mismatch DNA Hybridization properties Using DNA Hybridization Detection Sensor)

  • 김도균;권영수
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2003년도 추계학술대회 논문집 전기물성,응용부문
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    • pp.89-91
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    • 2003
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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Real-Time Detection of DNA Hybridization Assay by Using Evanescent Field Microscopy

  • Kim, Do-Kyun;Choi, Yong-Sung;Murakami, Yuji;Tamiya, Eiichi;Kwon, Young-Soo
    • KIEE International Transactions on Electrophysics and Applications
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    • 제11C권3호
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    • pp.85-90
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    • 2001
  • The determination of DNA hybridization reaction can apply the molecular biology research, clinic diagnostics, bioengineering, environment monitoring, food science and other application area. So, the improvement of DNA detection system is very important for the determination of this hybridization reaction. In this study, we report the characterization of the probe and target oligonucleotide hybridization reaction using the evanescent field microscopy. First, we have fabricated DNA chip microarray. The particles which were immobilized oligonucleotides were arranged by the random fluidic self-assembly on the pattern chips, using hydrophobic interaction. Second, we have detected DNA hybridization reaction using evanescent field microscopy. The 5'-biotinylated probe oligonucleotides were immobilized on the surface of DNA chip microarray and the hybridization reaction with the Rhodamine conjugated target oligonucleotide was excited fluorescence generated on the evanescent field microscopy. In the foundation of this result, we could be employed as the basis of a probe olidonucleotide, capable of detecting the target oligonucleotide and monitoring it in a large analyte concentration range and various mismatching condition.

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

  • 정우성;이웅희;백세환
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2001년도 추계학술발표대회
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    • pp.235-236
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    • 2001
  • 유전자 분석 기술은 biomedical analysis를 위해 일반적으로 고체 상에 고정화된 DNA 분자를 이용한다. 이 센서의 검출능력은 주로 capture probe의 서열뿐만 아니라 oligonucleotide의 고체 상에 고정화 방법에 달려있다. 본 연구에서는 glass 표면에 DNA 분자를 고정화시키는 방법과 PCR product를 정제하지 않고 직접 검출할 수 있는 DNA probe assay 방법을 개발하였다.

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Array-based Nano-amplification Technique Was Applied in Detection of Hepatitis E Virus

  • Liu, Hui-Hui;Cao, Xuan;Yang, Yong;Liu, Ming-Gui;Wang, Ye-Fu
    • BMB Reports
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    • 제39권3호
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    • pp.247-252
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    • 2006
  • A rapid method for the detection of Hepatitis E Virus (HEV) was developed by utilizing nano-gold labeled oligonucleotide probes, silver stain enhancement and the microarray technique. The 5'-end -$NH_2$ modified oligonucleotide probes were immobilized on the surface of the chip base as the capture probe. The detection probe was made of the 3'-end -SH modified oligonucleotide probe and nano-gold colloid. The optimal concentrations of these two probes were determined. To test the detection sensitivity and specificity of this technique, a conservative fragment of the virus RNA was amplified by the RT-PCR/PCR one step amplification. The cDNA was hybridized with the capture probes and the detection probes on microarray. The detection signal was amplified by silver stain enhancement and could be identified by naked eyes. 100 fM of amplicon could be detected out on the microarray. As the results, preparation of nano-gold was improved and faster. Development time also was shortened to 2 min. Thus, considering high efficiency, low cost, good specificity and high sensitivity, this technique is alternative for the detection of HEV.

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

  • 정우성;백세환
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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    • pp.709-710
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    • 2000
  • 유전자센서 기술은 biomedical analysis를 위해 일반적으로 고체 상에 고정화된 DNA 분자를 이용한다. 이 센서의 검출능력은 주로 capture probe의 서열뿐만 아니라 oligonucleotide의 고체 상에 고정화 방법에 달려있다. 본 연구에서는 glass 표면에 DNA 분자를 고정화시키는 두 가지 다른 방법을 비교하였고 유전자센서의 구성에 대해 검토하였다.

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DNA 측정용 SAW 센서의 주파수 증대에 의한 감도향상 (Improvement in Sensitivity by Increasing the Frequency of SAW Sensors for DNA Detection)

  • 사공정열;김재호;이수석;노용래
    • 한국음향학회지
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    • 제26권1호
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    • pp.42-47
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    • 2007
  • 본 논문에서는 probe DNA의 고정화 및 Probe DNA와 target DNA의 혼성화 반응을 감지할 수 있는 DNA 측정용 고주파 SAW 센서의 주파수 증대에 따른 감도향상에 대해 연구하였다. 센서는 $36^{\circ}$ YX $LiTaO_3$ 압전 단결정 기판위에 Au 박막이 증착된 측정채널 (sensing channel)과 기준채널 (reference channel)로 구성되며 200MHz에서 발진되는 이중 지연선 형태로 제작되었다. 또한 SAW 센서의 감지 미케니즘의 최적화를 위해 SAW 센서의 Au 지연선상의 Probe DNA의 최적 고정화 반응농도와 target DNA의 최적 혼성화 반응농도를 결정하였으며, 디지털 시린지 펌프시스템을 구성하여 실험자에 따른 오차를 최소화하였다. 측정채널의 Au 박막 지연선상에 probe DNA를 고정화시킨 후 target DNA를 주입하면, DNA의 혼성화 반응이 일어나며 Au 지연선상의 질량이 변하게 된다. 따라서 질량하중 효과에 대한 센서의 주파수 변화를 측정하였다. 개발된 센서는 최대 0.066ng/ml/Hz의 민감도를 가지며 질량하중 효과에 대한 안정적인 주파수 변화를 나타내었다.

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

  • 허영준;박유근;노용래
    • 한국음향학회지
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    • 제24권3호
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    • pp.160-165
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    • 2005
  • 본 연구에서는 DNA의 고정화 및 DNA 혼성화 반응을 감지할 수 있는 SH형 SAW 센서를 개발하였다. 고정화 및 혼성화 반응에 사용된 탐침 DNA 및 표적 DNA는 상보적 결합이 일어날 수 있는 염기서열을 가진 15-mer의 올리고뉴클레 오티드를 사용하였다. SH형 SAW 센서는 압전 단결정 $36^{\circ}\;YX\;LiTaO_3$를 사용하여 100 MHz로 발진되는 이중 지연선 형태로 제작하였다. 제작된 센서는 Au가 증착된 박막위에 고정화된 탐침 DNA와 표적 DNA와의 혼성화 반응을 시키고 난 후 센서의 주파수 변화를 측정하였으며, DNA 고정화 및 혼성화 반응은 pH 7.4의 PBS 완충용액상에서 수행하였다. 개발된 SH형 SAW센서는 $1.55 {\cal}ng/{\cal}ml/Hz$의 민감도를 가지며, DNA 혼성화 특성에 기인한 질량하중 효과에 따른 안정적인 주파수 변화를 나타내었다.