• Title/Summary/Keyword: Continuous Flow PCR

검색결과 11건 처리시간 0.025초

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

  • 정승룡;김준혁;이인제;강치중;김용상
    • 전기학회논문지
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    • 제56권3호
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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.

마이크로 칩(ATMEGA128)을 이용한 PCR의 온도제어 (Using the micro-chip (ATMEGA128) for controling of temperature)

  • 임기태;박민호;이한별;양기훈;이병성;한다운;김용상
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2011년도 제42회 하계학술대회
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    • pp.59-60
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    • 2011
  • 본 논문에서는 기존의 PCR 장비가 가지고 있는 낮은 경제성, 장비의 대형화, 긴 분석 시간 등과 같은 단점을 해결하기 위하여 ATMEGA128 마이크로 칩을 사용 continuous-flow PCR 칩의 온도를 제어 하였다. Polydimethylsiloxane (PDMS)와 산화 인듐-주석(Indium tin-oxide, ITO) 유리 기판을 사용하여 continuous-flow PCR 칩을 제작하였고 PDMS를 주조 하여 마이크로 채널을 형성하였다. 또한 유리 기판위에 ITO 전극을 패터닝하여 마이크로 히터를 제작하였다. 이 결과 continuous-flow PCR 칩에서 빠르고 정확한 온도 제어를 통한 DNA 중합 효소 연쇄반응 결과를 얻을 수 있었다.

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연속류형 Micro-PCR 시스템의 설계 (Design of Continuous-flow Micro-PCR System)

  • 김덕종;김재윤;박상진;허필우;윤의수
    • 유체기계공업학회:학술대회논문집
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    • 유체기계공업학회 2003년도 유체기계 연구개발 발표회 논문집
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    • pp.115-120
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    • 2003
  • In this work, a continuous-flow micro-PCR system is systematically designed. From the numerical simulation based on the finite volume method, adapting oneself to a new environmental temperature without an external temperature controller is shown to be possible and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. In addition, appropriate geometry of a heat sink for the cooler is determined by using a compact modeling method, the porous medium approach.

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연속식 PCR과 BCR에서 기$cdot$액 접촉 방법 및 주입 오존 유량에 따른 페놀 폐수의 오존 처리특성 (The Characteristics of the Ozonation of the Phenol Wastewater in the Continuous PCR and BCR)

  • 김용대;안재동;이준
    • 한국환경보건학회지
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    • 제22권1호
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    • pp.57-64
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    • 1996
  • The main objectives of this research program were to study the ozonation characteristics of phenol wastewater in the continuous packed colamn reactor(PCR) and the bubble column reactor (BCR) using ozone and to provide the fundamentals of ozonizing the phenol wastewater. Among various influencing factors that affect on phenol decomposition through the oxidation by ozone, contacting method, and ozone flow rate were chosen as reaction parameters. The results were obtained from two different types of contacting methods where the countercurrent flow was more efficient than the cocurrent flow in both the phenol removal efficiency and the ozone utilization efficiency. Furthermore, PCR showed the phenol removal efficiency 1.6 to 3% higher than that of BCR in both contacting methods, as well as the ozone utilization efficiency, suggesting that the countercurrent flow is more efficient than the cocurrent flow. The phenol removal efficiency and the ozone utilization efficiency were reduced in both reactors as the influent ozone flow rate increased. Upon varing flow rate from 0.5l/min to 2.0 l/min by 0.5 l/min, the phenol removal efficiency was reduced approximately from 8.5% to 10.5% and the ozone utilization efficiency was reduced approximately from 6% to 8% in both reactors. The performance of PCR was superior to that of BCR in the aspects of phenol removal and ozone utilization efficiency.

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연속흐름 중합효소연쇄반응칩 제작을 위한 인듐 산화막 전극의 특성분석 (Characteristics of Indium-Tin-Oxide electrode for continuous-flow PCR chip)

  • 정승룡;이인제;김준혁;김한수;김재완;최영진;강치중;김용상
    • 대한전기학회:학술대회논문집
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    • 대한전기학회 2006년도 제37회 하계학술대회 논문집 C
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    • pp.1386-1387
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    • 2006
  • PDMS와 ITO 유리를 이용하여 continuous-flow PCR chip을 제작하였다. PDMS를 이용하여 microchannel을 형성하여 주었고, ITO electrode를 heater와 sensor로 사용하기 위하여 반도체 공정을 통해 패턴을 형성하였다. microchannel내에 흐르는 시료의 온도를 제어하기 위하여 heater와 sensor를 calibration을 하였다. ITO heater는 인가된 전압에 대해 매우 선형적인 발열을 하였으며, ITO sensor는 온도에 대해 선형적인 저항 변화를 나타낸 바, 그 결과 continuous-flow PCR chip의 정확한 온도 제어가 가능하였다.

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랩온어칩을 위한 중합효소 연쇄반응 칩의 열설계 (Thermal Design of PCR Chip for LOC)

  • 김덕종;김재윤;박상진;허필우;윤의수
    • 연구논문집
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    • 통권33호
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    • pp.17-25
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    • 2003
  • In this work, thermal design of a PCR chip for LOC is systematically conducted. From the numerical simulation of a PCR chip based on the finite volume method, how to control the average temperature of a PCR chip and the temperature difference between the denaturation zone and the annealing zone is presented. The average temperature is shown to be controlled by adjusting heat input and a cooler as well as a heater is shown to be necessary to obtain three individual temperature zones for polymerase chain reaction. To reduce the time required, a heat sink for the cooler is not included in the calculation domain for the PCR chip and heat sink design is conducted separately by using a compact modeling method, the porous medium approach.

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Anti-obese and Blood Flow Improvement Activities of Ginseng Berry on the 45%Kcal High Fat Diet Supplied Mouse

  • Lee, Sol;Lee, Hae-Jeung;Chun, Yoon-Seok;Seol, Du-jin;Kim, Jong-Kyu;Ku, Sae-Kwang;Lee, Young-Joon
    • 대한예방한의학회지
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    • 제22권1호
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    • pp.107-127
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    • 2018
  • Objectives : The present study investigated the anti-obese and blood flow improvement activities of aqueous extracts of ginseng berry (GBe) on the mild diabetic obese mice as compared with metformin. Methods : After end of 56 days of continuous oral administrations of GBe 150, 100 and 50 mg/kg, or metformin 250 mg/kg, anti-obese and blood flow improvement effects - the changes of body weights, body and abdominal fat density by in live dual-energy x-ray absorptionmetry (DEXA), tail bleeding time, prothrombin time (PT), activated partial thromboplastin time (aPTT), serum total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDL) and high density lipoprotein (HDL) levels, aorta and serum cyclic guanosine monophosphate (cGMP), nitric oxide (NO) and endothelin (ET)-1 levels, aorta phosphorylated PI3K (pPI3K), phosphorylated Akt (pAkt) and phosphorylated p38 MAPK (pp38 MAPK) levels were systemically analyzed. In addition, aorta vascular dilation and constriction related gene mRNA expressions - PI3K, Akt, eNOS, p38 MAPK and ET-1 were also analyzed by realtime RT-PCR. Results : The obesity and related blood flow impairment, induced by 84 days of continuous HFD supply, were significantly inhibited by 56 days of continuous oral treatment of GBe 150, 100 and 50mg/kg, dose-dependently, and they also dramatically normalized the changes of the aorta vascular dilation and constriction related gene mRNA expressions, also dose-dependently. Especially, GBe 150 mg/kg constantly showed favorable inhibitory activities against type II diabetes related obesity and vascular disorders through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities, as comparable to those of metformin 250 mg/kg in HFD mice. Conclusion : By assessing the key parameters for anti-obese and blood flow improvement activities on the HFD-induced mild diabetic obese mice, the present work demonstrated that GBe 150, 100 and 50 mg/kg showed favorable anti-obese and blood flow improvement effects in HFD-induced type II diabetic mice, through PI3K/Akt pathway and p38 MAPK mediated cGMP, NO and ET-1 expression modulatory activities.

Polydimethylsiloxane 기반 미세유체시스템의 음향열적 가열 및 응용 (Acoustothermal Heating of Polydimethylsiloxane Microfluidic Systems and its Applications)

  • 성형진;하병항;박진수;굴람 데스트기르;정진호
    • 한국가시화정보학회지
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    • 제14권1호
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    • pp.57-61
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    • 2016
  • We report a finding of fast(exceeding 2,000 K/s) heating of polydimethylsiloxane(PDMS), one of the most commonly-used microchannel materials, under cyclic loadings at high(~MHz) frequencies. A microheater was created based on the finding. The heating mechanism utilized vibration damping of sound waves, which were generated and precisely manipulated using a conventional surface acoustic wave(SAW) microfluidic system, in PDMS. The penetration depths were measured to range from $210{\mu}m$ to $1290{\mu}m$, enough to cover most microchannel heights in microfluidic systems. The energy conversion efficiency was SAW frequency-dependent and measured to be the highest at around 30 MHz. Independent actuation of each interdigital transducer(IDT) enabled independent manipulation of SAWs, permitting spatiotemporal control of temperature on the microchip. All the advantages of this microheater facilitated a two-step continuous flow polymerase chain reaction(CFPCR) to achieve the billion-fold amplification of a 134 bp DNA amplicon in less than 3 min. In addition, a technique was developed for establishing dynamic free-form temperature gradients(TGs) in PDMS as well as in gases in contact with the PDMS.

Effects of high Cell Density on growth-Associated Monoclonal Antibody Production by Hybridoma T0405 Cells Immobilized in Macroporous Cellulose carriers

  • Hideki Mochoda;Wang, Pi-Chao;Fr Jr. Nayve;Ryuji Sato;Minoru Harige;Nakao Nomura;Masatoshi Matsumura
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제5권2호
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    • pp.110-117
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    • 2000
  • Relationship between monoclonal antibody (MAb) productivity and growth rate, and effects of high cell density on MAb production rate increased with increasing specifis growth rate in both suspended and immobilized continuous cultures indicate a positively growth-associated relationship between MAb productivity and growth rate. moreover, the specific production rate was higher in the immobilized cell culture than that in suspended one at all dilution rates. In order to clarify these phenomana, MAb mPNA experession and cell cycle distribution were investigated in bacth cultures with immobilized cells and suspended cells. RT-PCR was used for observation of MAb mRNA expression and a two-color bromodeoxyuridine (BrdU)/propidium iodide (PI) flow cytometry method for determination of cell cycle distribution. The results revealed that MAb nRNA expression until dead phase, which was longer than in suspended cell. The cell cycle distribution patterns were observed almost the same for both immobilized and suspended cells. Such results may imply that a high cell density state has positive influence on the mRNA expression and on growth-associated Mab productivity of T0405 cells.

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고정화 혼합효소를 이용한 새로운 구조의 올리고당 생산 및 특성 연구 (Production and Characterization of New Structured-Oligosaccharides from Immobilized Mixed-enzyme Reactor)

  • 이인수;김도만;허수진;김문수;이기영
    • KSBB Journal
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    • 제15권1호
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    • pp.88-95
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    • 2000
  • 5%(w/v)의 설탕과 2.5%(w/v)의 녹말로 구성된 혼합기질용액을 1.5 mL/min으로 공급하였을 때 dextransucrase와 $\alpha$-amylase의 혼합효소를 함께 고정화한 비드를 이용한 방법이 두 효소를 띠로 고정화한 비드를 혼합한 방법보다 올리고당 수율이 충전형 반응기에서는 12%, 교반형 반응기에서는 11% 높았다. 혼합효소를 함께 고정화한 비드를 200시간 이상 사용하였을 경우 교반식 반응기에서는 파괴되었으나, 충전형 반응기에서는 안전하였다. 따라서 올리고당을 연속적으로 생산하는데는 충전형 반응기가 더 적합하였다. 충치균인 S. mutans를 새로운 구조의 올리고당이 함유된 배치에서 배양하였을 때 pH가 5이하로 떨어지지 않았다. 식중독균인 S. aureus를 새로운 구조의 올리고당이 첨가된 배치에서 배양한 결과, glucose와 설탕이 첨가된 배치보다 균 성장정도가 현저히 떨어졌다.

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