• Title/Summary/Keyword: cDNA chip

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Characteristics of Indium-Tin-Oxide Electrode for Continuous-flow PCR Chip (연속흐름 중합효소연쇄반응칩 제작을 위한 인듐 산화막 전극의 특성분석)

  • Joung, Seung-Ryong;Kim, Jun-Hyeok;Yi, In-Je;Kang, C.J.;Kim, Yong-Sang
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.3
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    • pp.561-565
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    • 2007
  • We propose glass and PDMS (polydimethylsiloxane) chips for DNA amplification with continuous-flow PCR (polymerase chain reaction). The PDMS microchannel was fabricated using a negative molding method for sample injection. Three heaters and sensors of ITO (indium-tin-oxide) thin films were fabricated on glass chip. ITO heaters and sensors were calibrated accurately for the temperature control of the liquid flow. ITO heater generated stable heat versus applied power. ITO sensor resistance was changed linearly versus temperature increase as a RTD (resistance temperature detector) sensor. As a result, we enable precision temperature control of continuous-flow PCR chip. Using the continuous-flow PCR chip DNA plasmid pKS-GFP 720 bp was successfully amplified.

A Method for Evaluation of the Quality of DNA Microarray Spots

  • Zhang, Bao;Ma, Wen-Li;Hu, Zi-You;Shi, Rong;Song, Yan-Bin;Zheng, Wen-Ling
    • BMB Reports
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    • v.35 no.5
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    • pp.532-535
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    • 2002
  • To establish a method to evaluate the quality of the printed microarray and DNA fragments' immobilization. The target gene fragments that were made with the restriction display PCR (RD-PCR) technique were printed on a superamine modified glass slide, then immobilized with UV cross-linking and heat. This chip was hybridized with universal primers that were labeled with cy3-dUTP, as well as cDNA that was labeled with cy3-dCTP, as the conventional protocol. Most of the target gene fragments on the chip showed positive signals, but the negative control showed no signal, and vice versa. We established a method that enables an effective evaluation of the quality of the microarrays.

The Effects of Echinacea Extract on the Gene Expression of Monocytes and Monocyte-derived Dendritic Cells (Echinacea 추출물이 단구와 단구유래 수지상세포의 유전자발현에 미치는 효과)

  • Park, Jun Eun;Choi, Kang Duk;Kim, Sung Hwan;Hahm, Dae-Hyun;Seo, Jong Jin
    • Clinical and Experimental Pediatrics
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    • v.48 no.7
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    • pp.779-788
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    • 2005
  • Purpose : Echinacea, a traditional plant medicine has been used as immune-stimulant. Recent studies have revealed that extract of Echinacea has immunostimulatory effects on human blood mononuclear cells. This study was designed for the purpose of screening the genes associated with immunologic effects of Echinacea on monocytes and dendritic cells using a cDNA microarray chip. Methods : $CD14^+$ monocyte cells were cultured for one day with Echinacea extract(final concentration : $50{\mu}g/mL$) in experiment 1, but were cultured without Echinacea in experiment 2. The gene expression of these cultured monocytes was analyzed using the cDNA microarray chip. Dendritic cells produced from $CD14^+$ monocyte were cultured for five days with GM-CSF and IL-4, and then cultured for one day with Echinacea in experiment 3, but were done without Echinacea in experiment 4. Results : In experiments 1 and 2, there were 17 significantly expressed genes with average expression ratios above 2.5, including interferon gamma-inducible protein 30(IFI 30), CDC(cell-division-cylcle)-like kinase 2(CLK 2), syndecan binding protein(syntenin), superoxide dismutase 2, etc. In experiments 3 and 4, there were 24 gene, with significantly expressed genes were 24 genes, which were insulin-like growth factor 2(somatomedin A), methyl-CpG binding domain protein 3, IFI 30, small inducible cytokine subfamily A, member 22, etc. The genes encoding CD44, IFI 30, mannose receptor C type 1(MRC 1), chemokine receptor 7(CCR 7), CLK 2, syntenin and cytochrome C oxidase subunit VIII were significantly expressed in both monocytes and dendritic cells cultured with Echinacea. Conclusion : This study employed a cDNA microarray chip to elicit the immune-associated gene profile; the expression was enhanced by Echinacea in CD14+ monocytes and dendritic cells. Thus we laid the basis for the quantitative and functional analysis of genes induced by Echinacea in monocytes and monocyte-derived dendritic cells.

Study on Microbiochip for Buccal Cell Lysis and DNA Purification (상피세포 시료 전처리용 마이크로바이오칩에 관한 연구)

  • Ha, Seung-Mo;Cho, Woong;Ahn, Yoo-Min;Hwang, Seung-Yong
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.34 no.12
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    • pp.1785-1791
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    • 2010
  • This paper describes a separable microfluidic device fabricated with PDMS (polydimethylsiloxane) and glass. The device is used for sample preparation involving cell lysis and the DNA purification process. The cell lysis was performed for 2 min at $80^{\circ}C$ in a serpentine-type microreactor ($20 {\mu}l$) using a Au microheater that was integrated with a thermal microsensor on a glass substrate. The DNA that was mixed with other residual products during the cell lysis process was then filtered through a new filtration system composed of microbeads (diameter: $50 {\mu}m$) and PDMS pillars. Since the entire process (sample loading, cell lysis reaction, DNA purification, and sample extraction) was performed within 5 min in a microchip, we could reduce the sample preparation time in comparison with that for the conventional methods used in biochemistry laboratories. Finally, we verified the performance of the sample preparation chip by conducting PCR (polymerase chain reaction) analysis of the chip product.

Studies on Cellular Factors Responsible for 2,3,7,8-TCDD Resistency and Cellular Transformation (2,3,7,8-TCDD의 세포형질전환 및 내성획득에 관여하는 세포내 인자에 관한 연구)

  • Ryeom Tai-Kyung;Choi Young-Sill;Kim Ok-Hee;Kang Ho-Il
    • Environmental Mutagens and Carcinogens
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    • v.26 no.1
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    • pp.1-6
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    • 2006
  • To enhance our understanding of toxicity mediated through the pathway by which TCDD stimulates gene expression, we have investigated genes whose expressions are changed after treatment with TCDD and/or MNNG in human Chang liver cell. First, we treated with MNNG and TCDD for two weeks to transform human Chang liver cell. We obtained cell looks like to be transformed and compared the differential gene expression by using cDNA chip (Macrogen) which carrys genes related with signal transduction pathways, oncogenes and tumor suppressor genes, etc. We found that TCDD up- or down-regulated 203 and 111 genes including oncogenes and tumor suppressor genes in human Chang liver cell two fold or more, respectively. Second, we compared the differential gene expression after treatment with TCDD only by using cDNA chip (Superarray) which carrys genes related with cell cycle regulations, and found that TCDD up regulated genes related with cell proliferation as well as cell growth inhibition in human Chang liver cell two fold or more, respectively. These results suggest that toxicity induced by TCDD may reflect sustained alterations in the expression of many genes and that the changes reflect both direct and indirect effects of TCDD.

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Effects of Sinomenium acutum Extract on Body Weight Gains and the DNA Chip Expression of Obese Rats. (방기(防己)의 투여가 비만 유발 쥐의 생리기능과 DNA Chip을 통한 유전자 발현에 미치는 영향에 대한 연구)

  • Joh, Ho-Geun;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.4
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    • pp.41-55
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    • 2007
  • Purpose: This study is to examine anti-obesity effect and cytotoxicity of the long-term oral administration of Sinomenium acutum (Bang-gi, SA) Methods: Using diet-induced obesity C57BL/6 mouse model, anti-obesity effect and DNA chip expression and cytotoxicity of the long-term oral administration of this herbal extract were investigated. Results: The herbal extract treated groups were arrested in weight increment only when they were lodged together. Such effects were abolished when they kept individually. SA fed mice behaved very rudely and violently. On the basis of histological studies of liver tissues and also in vitro cytotoxicity tests of the liver and kidney cell lines, no significant toxicity was found by 14 weeks of SA treatments. However, we found significant changes in gene expression profile in SA treated group by micro-array analysis. In case of SA group, up-regulated genes were 1,213 and down-regulated ones were 2,558. Some of lipid metabolism related genes also significantly changed in both treatment groups. Conclusion: SA had effects of increasing the basal metabolic rate by stimulating the sympathetic nervous systems.

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Effects of the oral administration of Epedra Sinica Extract on suppression of body weight gains and the DNA chip expression of obese rats. (마황(麻黃)의 투여가 비만 유발 쥐의 생리기능과 DNA Chip을 통한 유전자 발현에 미치는 영향에 대한 연구)

  • Joh, Ho-Geun;Yang, Jeong-Min;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.20 no.3
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    • pp.65-80
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    • 2007
  • Purpose: This study is to examine anti-obesity effect and cytotoxicity of the long-term oral administration of Ephedra Sinica(Ma-hwang, ES) Methods: Using diet-induced obesity C57BL/6 mouse model, anti-obesity effect and DNA chip expression and cytotoxicity of the long-term oral administration of this herbal extract were investigated. Results: The herbal extract treated groups were arrested in weight increment only when they were lodged together. Such effects were abolished when they kept individually. ES fed mice behaved very rudely and violently. On the basis of histological studies of liver tissues and also in vitro cytotoxicity tests of the liver and kidney cell lines, no significant toxicity was found by 14 weeks of ES treatments. However, we found significant changes in gene expression profile in ES treated group by micro-array analysis. In case of ES group, up-regulated genes were 113 and down-regulated were 120. Some of lipid metabolism related genes also significantly changed in treatment groups. Conclusion: ES had effects of increasing the basal metabolic rate by stimulating the sympathetic nervous systems.

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PDMS/Glass Serpentine Microchannel Chip for PCR with Bubble Suppression in Sample Injection (시료주입시 기포발생이 억제된 반응조 형태의 중합효소연쇄반응용 PDMS/유리 바이오칩)

  • Cho Chul-Ho;Cho Woong;Hwang Seung-Yong;Ahn Yoo-Min
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.30 no.10 s.253
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    • pp.1261-1268
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    • 2006
  • This paper reports low-cost microreactor $(10{\mu}{\ell})$ biochip for the DNA PCR (polymerase chain reaction). The microbiochip $(20mm{\times}28mm)$ is a hybrid type which is composed of PDMS (polydimethylsiloxane) layer with serpentine micochannel $(360{\mu}m{\times}100{\mu}m)$ chamber and glass substrate integrated with microheater and thermal microsensor. Undesirable bubble is usually created during sample loading to PMDS-based microchip because of hydrophobic chip surface. Created bubbles interrupt stable biochemical reaction. We designed improved microreactor chamber using microfluidic simulation. The designed reactor has a coner-rounded serpentine channel architecture, which enables stable injection into hydrophobic surface using micropipette only. Reactor temperature needed to PCR reaction is controlled within ${\pm}0.5^{\circ}C$ by PID controller of LabVIEW software. It is experimentally confirmed that SRY gene PCR by the fabricated microreactor chip is performed for less than 54 min.

Design of an Inexpensive Heater using Chip Resistors for a Portable Real-time Microchip PCR System (저항소자를 이용한 휴대형 Real-time PCR 기기용 히터 제작)

  • Choi, Hyoung-jun;Kim, Jeong-tae;Koo, Chi-wan
    • Journal of IKEEE
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    • v.23 no.1
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    • pp.295-301
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    • 2019
  • A heater in a portable real-time polymerase chain reaction(PCR) system is one of the important factors for controlling the PCR thermocycle precisely. Since heaters are integrated on a small-sized PCR chip for rapid heating and fabricated by semiconductor processes, the cost of producing PCR chips is high. Here, we propose to use chip resistors as an inexpensive and accurate temperature control method. The temperature distribution was simulated using one or two chip resistors on a real-time PCR chip and the PCR chip with uniform temperature distribution was fabricated. The temperature rise and fall rates were $18^{\circ}C/s$ and $3^{\circ}C/s$, respectively.

Gene Expression Analysis Using cDNA Microarray Assay by Cervi Pantotricuhum Cornu Herbal Acupuncture (녹용약침액(鹿茸藥鍼液)의 DNA chip을 이용(利用)한 유전자(遺傳子) 발현(發顯) 분석(分析))

  • Han, Sang-won;Seo, Jung-chul;Lee, Yun-ho;Choi, Je-yong
    • Journal of Acupuncture Research
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    • v.20 no.3
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    • pp.34-44
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    • 2003
  • Objective : Bone homeostasis is maintained by balance of bone formation and resorption. Therefore, bone related diseases arose by disturbance of this balance between osteoblast and osteoclast activities. To develop a successful screening system the therapeutic components based on oriental medicine is essential to set up systematic approach for that purpose. The purpose of this study is to the know the gene expression using cDNA microarray assay. Methods : Cervi Pantotricuhum Cornu Herbal-acupuncture extract was prepared by boiling. human osteosarcoma cells(HOS) were treated with Cervi Pantotricuhum Cornu Herbal-acupuncture solution. Then mRNA was extracted and cDNA microarray assay was performed. Results : Human osteosarcoma cells(HOS) treated with Cervi Pantotricuhum Cornu Herbal-acupuncture solution($500{\mu}g/m{\ell}$) showed that thioredoxin, TAFII31 and two novel genes were increased. However many genes decreased their expression by Cervi Pantotricuhum Cornu Herbal-acupuncture. Conclusions : This type of approach will give a good chance to explore the favorable effects of Cervi Pantotricuhum Cornu Herbal-acupuncture. Further study is needed for investigating the effect of Cervi Pantotricuhum Cornu Herbal-acupuncture.

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