• Title/Summary/Keyword: immobilized oligonucleotide probe

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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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    • v.7 no.4
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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.

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

  • Kim, Do-Kyun;Kwon, Young-Soo
    • Proceedings of the KIEE Conference
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    • 2003.10a
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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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    • v.11C no.3
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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

  • 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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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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    • v.39 no.3
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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

  • 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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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.

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 $1.55{\cal}ng/{\cal}ml/Hz$.