• 제목/요약/키워드: Electrophoresis

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An Inexpensive System for Rapid and Accurate On-site Detection of Garlic-Infected Viruses by Agarose Gel Electrophoresis Followed by Array Assay

  • Kazuyoshi Furuta;Shusuke Kawakubo;Jun Sasaki;Chikara Masuta
    • The Plant Pathology Journal
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    • 제40권1호
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    • pp.40-47
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    • 2024
  • Garlic can be infected by a variety of viruses, but mixed infections with leek yellow stripe virus, onion yellow dwarf virus, and allexiviruses are the most damaging, so an easy, inexpensive on-site method to simultaneously detect at least these three viruses with a certain degree of accuracy is needed to produce virus-free plants. The most common laboratory method for diagnosis is multiplex reverse transcription polymerase chain reaction (RT-PCR). However, allexiviruses are highly diverse even within the same species, making it difficult to design universal PCR primers for all garlic-growing regions in the world. To solve this problem, we developed an inexpensive on-site detection system for the three garlic viruses that uses a commercial mobile PCR device and a compact electrophoresis system with a blue light. In this system, virus-specific bands generated by electrophoresis can be identified by eye in real time because the PCR products are labeled with a fluorescent dye, FITC. Because the electrophoresis step might eventually be replaced with a lateral flow assay (LFA), we also demonstrated that a uniplex LFA can be used for virus detection; however, multiplexing and a significant cost reduction are needed before it can be used for on-site detection.

전기영동법(電氣泳動法)에 의(依)한 대두(大豆) 근류균(根瘤菌) Rhizobium fredii와 Bradyrhizobium japonicum의 분류(分類) 및 동정(同定) (Identification of Rhizobium fredii and Bradyrhizobium japonicum by Polyacrylamide Gel Electrophoresis)

  • 윤한대;조무제;이계호
    • Applied Biological Chemistry
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    • 제30권2호
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    • pp.163-168
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    • 1987
  • 전보에서 보고한 Bradyrhizobium japonicum 9 균주와 Rhizobium fredii 7 균주의 균체 중의 단백질 Pattern의 차이를 일차원 및 이차원 전기영동법에 의하여 조사하였다. 일차원 전기영동법 (SDS-PAGE)에 의해서는 두 group의 균체에서 모두 52개의 band가 관찰되었고 그 중 6개의 main band 로써 group 간의 차이가 확인되었으며, 이차원 전기영동법(2D-PAGE)에 의한 두 group간에 단백질 구성은 Rhizobium fredii 에서는 단백질이 산성 쪽에, Bradyrhizobium japonicum 는 alkali성 쪽에 비교적 많이 분포되어 있었다. 또한 두 group 간의 균체 아미노산 조성을 조사한 결과 조선상의 뚜렷한 차이가 없었다. 분리된 근류균을 확인하는데 전기영동법이 유용하였으며, 일차원 전기영동법은 많은 균주를 신속하게 확인할 수 있었고 이차원 전기영동법은 해상력 및 분리된 단백질 spot를 분석하는데 용이하였다.

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팽이버섯의 핵형분석과 균주 사이의 핵형 다양성 (Electrophoretic karyotype of Flammulina velutipes and its variation among cultivars)

  • 이송희;이미경;김나리;이창윤;이현숙
    • 한국버섯학회지
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    • 제12권1호
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    • pp.63-66
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    • 2014
  • 한국에서 개발된 팽이버섯 균주 Fv 3-6과 일본에서 수집한 팽이버섯 균주 Fv 0-1, Fv 1-5 및 Fv 11-4의 핵형을 CHEF gel electrophoresis 방법으로 분석 하였다. 그 결과 4종류의 균주 모두 chromosome의 전체 길이가 달랐으며, chromosome의 개수 또는 특정 chromosome의 길이가 다른 것을 확인하였다. 특히 Fv 3-6의 경우에는 다른 3 종의 균주와 비교했을 때 Fv 0-1, Fv 11-4 보다는 2개의 chromosome이 더 존재하였고 Fv 1-5 보다는 1개의 chromosome이 더 존재하였으며 핵형패턴이 유사한 일본 수집 균주들과는 다른 핵형패턴을 나타내었다. 이러한 CHEF gel electrophoresis 방법은 품종간의 차이를 SSR이나 ITS 정보를 이용한 방법보다 더 정확하게 구분할 수 있을 것이라고 생각한다.

Capillary Zone Electrophoresis를 이용한 음이온 동시분석에 관한 연구 (Analysis of Inorganic Acids by Capillary Zone Electrophoresis)

  • 박성우;진광호;유재훈;김동환;서배석;김영상
    • 분석과학
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    • 제11권3호
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    • pp.213-221
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    • 1998
  • Bromide, chloride, fluoride, nitrite, nitrate, sulfite, sulfate, perchlorate, chlorate 및 chlorite가 포함된 시료를 capillary zone electrophoresis(CZE)와 indirect UV detection 방법으로 분리하였다. 음이온류의 분리는 running buffer로 5 mM ammonium dichromate, 10 mM ammonium acetate, 20 mM diethylenetriamine, 10% methanol(pH 9.3)을 사용하였으며 전압은 cathode에 15 kV를 공급하였다. 또한 최초로 CZE에서 음이온의 정량이 가능하도록 각 이온들에 대한 regression과 detection limit를 결정하였고, 이 분석방법은 톨루엔을 흡입한 사람의 뇨, 부검 혈액, 폐수 및 폭약 등 시료에 적용하였으며, 음이온의 함량을 ion chromatograph(IC)와 비교 검토하였다.

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보리품종 구분에 적합한 전기영동법과 효소 (Appropriate Electrophoresis Techniques and Isozymes to Identification of Barley Cultivars)

  • 손응룡;이용세;윤경은;하용웅
    • 한국작물학회지
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    • 제30권4호
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    • pp.405-411
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    • 1985
  • 본 실험은 보리품종의 구분에 가장 적합한 전기영동법과 효소를 구명하고자 6개 보리품종(알찬보리, 수원 216호, 조풍보리, 사천 6호, 향맥, 히노데하다가)의 종실 단백질을 7.5% polyacrylamide slab gel, 2-30% polyacrylamide porosity gradient tube gel, isoelectricfocusing(pH 4-9)과 starch gel을 사용 전기영동 후 protein band pattern과 esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase 및 leucine aminopeptidase의 동위효소 pattern을 관찰하였던 결과는 다음과 같았다. 1. 7.5% polyacrylamide slab gel에서 단백질 pattern과 2-30% polyacrylamide porosity gradient tube gel에서의 단백질과 esterase의 band pattern이 품종간 뚜렷한 차이를 보여 보리품종구분에 가장 적합하였다. 2. Acid phosphatase, malate dehydrogenase, glutamate dehydrogenase 및 leucine aminopeptidase 등의 동위효소 pattern은 모든 실험한 품종이 동일한 형태를 보여 품종구분에 이용하기에는 적합하지 않았다.

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A proposal of new electronics device;micro-total analysis system for capillary electrophoresis

  • Oshige, Seisho;Aoyama, T.;Kambe, J.;Nagashima, U.
    • 제어로봇시스템학회:학술대회논문집
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    • 제어로봇시스템학회 2004년도 ICCAS
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    • pp.579-581
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    • 2004
  • We wish to develop micro-total analysis system (TAS) on a chip, and to make a trial approach to solve the important problem that is to detect ions separated by the electric field. We propose an idea, which is as for rotational motions of dipolar ions, which are affected by the ion atmosphere in outer regions. This is a new kind of the ion-sensitive field effect transistor (ISFET). We wish to develop the ISFET chips, and give more effective, fast and sensitive, capillary electrophoresis is designed in near future.

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Gel Electrophoresis Analysis of the Hard Coronas of Human Serum Albumin on Silica Nanoparticles: Size Dependence of Corona Formation

  • Kim, Sung-Jong;Han, Sang Yun
    • Bulletin of the Korean Chemical Society
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    • 제35권9호
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    • pp.2621-2624
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    • 2014
  • The rapid and spontaneous adsorption of proteins on nanoparticle (NP) surfaces in biological fluids such as blood is an important phenomenon as it possibly determines "what the cells see" and, thus, the fates of NPs in living organisms. In order to quantitatively understand protein coronas at the molecular level, we investigated human serum albumin (HSA) coronas that were produced on silica NPs of 20 nm and 50 nm diameters using conventional gel electrophoresis. Analysis of the concentration dependence of protein adsorption showed that HSA coronas preferentially formed a monolayer on silica NPs and revealed the presence of hard protein coronas. HSA adsorption was clearly dependent on NP size, and this might be due to the different surface curvatures of NPs of different sizes.

DNA damage와 Apoptosis를 정량화하는 단세포전기영동법 (Single Cell Gel Electrophoresis (comet assay) to Detect DNA Damage and Apoptosis in Cell Level)

  • 류재천;김현주;서영록;김경란
    • 한국환경성돌연변이발암원학회지
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    • 제17권2호
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    • pp.71-77
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    • 1997
  • The single cell gel electrophoressis(SCGE) assay, also known as the comet assay, is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakage in mammalian cells. The SCGE or comet assay is a promising test for the detection of DNA damage and repair in individnal cells. It has widespread potential applications in DNA damage and repair studies, genotoxicity testing and biomonitoring. In this microgel electrophoresis technique, cells are embedded in agarose gel on microscope slides, iysed and electrophoresed under alkaline conditions. Cells with increased DNA damage display increased migration of DNA from the nucleus towards the anode. The length of DNA migration indicates the amount of DNA breakage in the cell. The comet assay is also capable of identifying apoptotic cells which contain highly fragmented DNA. Here we review the development of the SCGE assay, existing protocols for the detection and analysis of comets, the relevant underlying principles determining the behaviour of DNA and the potential applications of the technique.

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모세관 전기영동법을 이용한 제제중의 수용성 비타민 분석 (Analysis of Water-soluble Vitamins in Pharmaceutical Products by Capillary Electrophoresis)

  • 이종진;문동철;이공주
    • 약학회지
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    • 제38권1호
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    • pp.38-45
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    • 1994
  • An analytical method using capillary electrophoresis (CE) was developed for quantitation of water soluble vitamin contents in various vitamin products. The method includes the optimization of separation of 11 water soluble vitamins changing the micellar concentration and pH of running buffer, applied voltage and sample preparation. Best resolution was obtained with 25 mM phosphate buffer (pH=8.0) containing 50 mM sodium dodecyl sulphate (SDS) as micellar phase. At optimum condition, water soluble vitamins were determined in orange juice and vitamin products such as vitamin C pulvis, vitamin injection, coated multivitamin tablet. The quantitative analysis of water soluble vitamins with CE was suitable for quality control of pharmaceutical products with sound reproducibility.

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비스페놀 A 및 Diethylstilbestrol의 유전독성 평가를 위한 염색체이상, 자매염색분체교환, 소핵형성, 단일세포 겔 전기영동법의 활용 (Analysis of chromosome aberration, sister chromatid exchange, micronuclei and single cell gel electrophoresis in human lymphocytes exposed in vitro to Bisphenol A and Diethylstilbestrol)

  • 김병모;정해원
    • 한국환경성돌연변이발암원학회지
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    • 제21권2호
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    • pp.135-141
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    • 2001
  • Endocrine disruptors have been implicated in carcinogenesis in animal studies, but carcinogenetic effects on human remain controversial. In order to examine the genotoxicity of two common endocrine disruptors, Bisphenol A and Diethylstilbestrol, cytogenetic endpoints including chromosome aberration (CA), sister chromatid exchange (SCE), micronuclei (MN) analyses and DNA damage by single cell gel electrophoresis (SCGE) were assessed. The effects of Bisphenol A and Diethylstilbestrol on the frequencies of CA and MN were increased in a dose-dependent manner and that of Bispheol A was more significant by Kendall'$\tau$test. Bisphenol A and Diethylstilbestrol also increased the frequency of SCE. Bisphenol A and Diethylstilbestrol induced DNA damage in a dose-dependent manner and the DNA damage induced by Diethylstilbestrol in human blood lymphocytes was more significant.

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