• Title/Summary/Keyword: Indicator-free DNA

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Hybridization by an Electrical Force and Electrochemical Genome Detection Using an Indicator-free DNA on a Microelectrode-array DNA Chip

  • Choi, Yong-Sung;Lee, Kyung-Sup;Park, Dae-Hee
    • Bulletin of the Korean Chemical Society
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    • v.26 no.3
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    • pp.379-383
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    • 2005
  • This research aims to develop DNA chip array without an indicator. We fabricated microelectrode array by photolithography technology. Several DNA probes were immobilized on an electrode. Then, indicator-free target DNA was hybridized by an electrical force and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between DNA probe and mismatched DNA in an anodic peak. Immobilization of probe DNA and hybridization of target DNA could be confirmed by fluorescent. This indicator-free DNA chip microarray resulted in the sequence-specific detection of the target DNA quantitatively ranging from $10^{-18}\;M\;to\;10^{-5}$ M in the buffer solution. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

Indicator-free DNA Chip Array Using an Electrochemical System

  • Park, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • v.4C no.4
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    • pp.133-136
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    • 2004
  • This research aims to develop a DNA chip array without an indicator. We fabricated a microelectrode array through photolithography technology. Several DNA probes were immobilized on an electrode. Then, target DNA was hybridized and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between the DNA probe and mismatched DNA in an anodic peak. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

Development of New DNA Chip and Genome Detection Using an Indicator-free Target DNA (비수식화 DNA를 이용한 유전자 검출 및 새로운 DNA칩의 개발)

  • Park, Yong-Sung;Park, Dae-Hee;Kwon, Young-Soo;Tomoji Kawai
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.52 no.8
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    • pp.365-370
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    • 2003
  • This research aims to develop an indicator-free DNA chip using micro-fabrication technology. At first, we fabricated a DNA microarray by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the gold electrodes. Then indicator-free target DNA was hybridized by an electrical force and measured electrochemically in potassium ferricyanide solution. Redox peak of cyclic-voltammogram showed a difference between target DNA and mismatched DNA in an anodic peak current. Therefore, it is able to detect various genes electrochemically after immobilization of various probe DNAs and hybridization of indicator-free DNA on the electrodes simultaneously It suggested that this DNA chip could recognize the sequence specific genes.

SNP (Single Nucleotide Polymorphism) Detection Using Indicator-free DNA (비수식화 DNA를 이용한 SNP의 검출)

  • Choi, Yong-Sung;Park, Dae-Hee;Kwon, Young-Soo
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2003.11a
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    • pp.224-226
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    • 2003
  • In this paper, we succeeded SNP discrimination of DNA hybridization on microarray using new electrochemical system. Using the electrochemical method with a label-free DNA has Performed DNA chip microarray. This method is based on redox of an electrochemical ligand. We developed scanning system with high performance.

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SNP Detection Using Indicator-free DNA Chip (비수식화 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Moon, Jong-Dae;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.06a
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    • pp.410-411
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    • 2006
  • High throughput analysis using a DNA chip microarray is powerful tool in the post genome era. Less labor-intensive and lower cost-performance is required. Thus, this paper aims to develop the multi-channel type label-free DNA chip and detect SNP (Single nucleotide polymorphisms). At first, we fabricated a high integrated type DNA chip array by lithography technology. Various probe DNAs were immobilized on the microelectrode array. We succeeded to discriminate of DNA hybridization between target DNA and mismatched DNA on microarray after immobilization of a various probe DNA and hybridization of label-free target DNA on. the electrodes simultaneously. This method is based on redox of an electrochemical ligand.

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Integrated Type DNA Chip Array and Gene Detection Using an Indicator-free DNA (집적형 DNA칩 어레이 및 비수식화 DNA를 이용한 유전자 검출)

  • Choi, Yong-Sung;Lee, Kyung-Sup
    • Proceedings of the KIEE Conference
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    • 2006.07c
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    • pp.1322-1323
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    • 2006
  • This research aims to develop the multiple channel electrochemical DNA chip that has the above characteristic and be able to solve the problems. At first, we fabricated a high integration type DNA chip array by lithography technology. It is able to detect a plural genes electrochemically after immobilization of a plural probe DNA and hybridization of non-labeling target DNA on the electrodes simultaneously.

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Development of Next Generation Biochip Using Indicator-free DNA (비수식화 DNA를 이용한 차세대형 바이오칩의 개발)

  • Choi, Yong-Sung;Moon, Jong-Dae;Lee, Kyung-Sup
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2006.05a
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    • pp.71-73
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    • 2006
  • This research aims to develop a multiple channel electrochemical DNA chip using micro- fabrication technology. At first, we fabricated a high integrated type DNA chip array by lithography technology. Several probe DNAs consisting of thiol group at their 5-end were immobilized on the gold electrodes. Then target DNAs were hybridized by an electrical force. Redox peak of cyclic-voltammogram showed a difference between target DNA and mismatched DNA in the anodic peak current. Therefore. it is able to detect a various genes electrochemically after immobilization of a various probe DNA and hybridization of label-free DNA on the electrodes simultaneously. It suggested that this DNA chip could recognize the sequence specific genes.

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Increased Free Circulating DNA Integrity Index as a Serum Biomarker in Patients with Colorectal Carcinoma

  • El-Gayar, Dina;El-Abd, Nevine;Hassan, Noha;Ali, Reem
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.939-944
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    • 2016
  • Background: Cell-free DNA circulating in blood is a candidate biomarker for malignant tumors. Unlike uniformly truncated DNA released from apoptotic non diseased cells, DNA released from necrotic cancer cells varies in size. Objectives: To measure the DNA integrity index in serum and the absolute DNA concentration to assess their clinical utility as potential serum biomarkers for colorectal carcinoma (CRC) compared to CEA and CA19-9. Materials and Methods: Fifty patients with CRC, 10 with benign colonic polyps and 20 healthy sex and age matched volunteers, were investigated by real time PCR of ALU repeats (ALU q-PCR) using two sets of primers (115 and 247 bp) amplifying different lengths of DNA fragments. The DNA integrity index was calculated as the ratio of q-PCR results of ALU 247/ALU 115bp. Results: Serum DNA integrity was statistically significantly higher in CRC patients compared to the benign and control groups (p<0.001). ROC curves for differentiating CRC patients from normal controls and benign groups had areas under curves of 0.90 and 0.85 respectively. Conclusions: The DNA integrity index is superior to the absolute DNA concentration as a potential serum biomarker for screening and diagnosis of CRC. It may also serve as an indicator for monitoring the progression of CRC patients. Combining CEA and CA19-9 with either of the genetic markers studied is better than either of them alone.

DNA Damage of Lymphocytes in Volunteers after 4 hours Use of Mobile Phone (일부 자원자들의 이동전화 4시간 연속 사용 후 림프구 DNA 손상 평가)

  • Ji, Seon-Mi;Oh, Eun-Ha;Sul, Dong-Geun;Choi, Jae-Wook;Park, Hee-Chan;Lee, Eun-Il
    • Journal of Preventive Medicine and Public Health
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    • v.37 no.4
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    • pp.373-380
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    • 2004
  • Objectives : There has been gradually increasing concern about the adverse health effects of electromagnetic radiation originating from cell phones which are widely used in modern life. Cell phone radiation may affect human health by increasing free radicals of human blood cells. This study has been designed to identify DNA damage of blood cells by electromagnetic radiation caused by cell phone use. Methods : This study investigated the health effect of acute exposure to commercially available cell phones on certain parameters such as an indicator of DNA damage for 14 healthy adult volunteers. Each volunteer during the experiment talked over the cell phone with the keypad facing the right side of the face for 4 hours. The single cell gel electrophoresis assay (Comet assay), which is very sensitive in detecting the presence of DNA strand-breaks and alkali-labile damage in individual cells, was used to assess peripheral blood cells (T-cells, B-cells, granulocytes) from volunteers before and after exposure to cell phone radiation. The parameters of Comet assay measured were Olive Tail Moment and Tail DNA %. Results : The Olive Tail Moment of B-cells and granulocytes and Tail DNA % of B-cells and granulocytes were increased by a statistically significant extent after 4-hour use of a cell phone compared with controls. Conclusion : It is concluded that cell phone radiation caused the DNA damage during the 4 hours of experimental condition. Nonetheless, this study suggested that cell phone use may increase DNA damage by electromagnetic radiation and other contributing factors.

Evaluation of Digital PCR as a Technique for Monitoring Acute Rejection in Kidney Transplantation

  • Lee, Hyeseon;Park, Young-Mi;We, Yu-Mee;Han, Duck Jong;Seo, Jung-Woo;Moon, Haena;Lee, Yu-Ho;Kim, Yang-Gyun;Moon, Ju-Young;Lee, Sang-Ho;Lee, Jong-Keuk
    • Genomics & Informatics
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    • v.15 no.1
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    • pp.2-10
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    • 2017
  • Early detection and proper management of kidney rejection are crucial for the long-term health of a transplant recipient. Recipients are normally monitored by serum creatinine measurement and sometimes with graft biopsies. Donor-derived cell-free deoxyribonucleic acid (cfDNA) in the recipient's plasma and/or urine may be a better indicator of acute rejection. We evaluated digital PCR (dPCR) as a system for monitoring graft status using single nucleotide polymorphism (SNP)-based detection of donor DNA in plasma or urine. We compared the detection abilities of the QX200, RainDrop, and QuantStudio 3D dPCR systems. The QX200 was the most accurate and sensitive. Plasma and/or urine samples were isolated from 34 kidney recipients at multiple time points after transplantation, and analyzed by dPCR using the QX200. We found that donor DNA was almost undetectable in plasma DNA samples, whereas a high percentage of donor DNA was measured in urine DNA samples, indicating that urine is a good source of cfDNA for patient monitoring. We found that at least 24% of the highly polymorphic SNPs used to identify individuals could also identify donor cfDNA in transplant patient samples. Our results further showed that autosomal, sex-specific, and mitochondrial SNPs were suitable markers for identifying donor cfDNA. Finally, we found that donor-derived cfDNA measurement by dPCR was not sufficient to predict a patient's clinical condition. Our results indicate that donor-derived cfDNA is not an accurate predictor of kidney status in kidney transplant patients.