• Title/Summary/Keyword: 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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    • v.40 no.1
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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.

Identification of Rhizobium fredii and Bradyrhizobium japonicum by Polyacrylamide Gel Electrophoresis (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))

  • Yun, Han-Dae;Cho, Moo-Je;Lee, Ke-Ho
    • Applied Biological Chemistry
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    • v.30 no.2
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    • pp.163-168
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    • 1987
  • A method, based upon the separation of cellular proteins by one-and two-dimensional electrophoresis was used for distinguishing butween Bradyrhizobium japonicum strains and Rhizobium fredii strains. Significant differences in protein pattern of one-dimensional SDS-PAGE vs-ere observed between Rhizobium fredii strains and Bradyrhizobium japonicum strains. The differences in six distinct main lands were observed among total 52 kinds of protein bands. Furthermore, the distribution of proteins in two groups by two-dimensional polyacrylamide gel electrophoresis was very different. The majority of visible proteins of Rhizobium fredii were acidic, whereas those of Bradyrhizobium japonicum were basic. In addition, amino acid composition was analyzed to detect the differences between two groups. No significant differences in amino acid composition were observed between Bradyrhizobium japonicum strains and Rhizobium fredii strains. The results indicate that one-and two-dimensional polyacrylamide gel electrophoresis were useful for identifying rhizobia isolates. One-dimensional SDS-PAGE of rhizobia proteins provided a rapid method for screening a large number of isolates, whereas two-dimensional electrophoresis was more of resolution and easiness for analyzing protein spots.

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

  • Lee, Song Hee;Lee, Mi-Kyoung;Kim, Na-Ri;Lee, Chang-Yun;Lee, Hyun-Sook
    • Journal of Mushroom
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    • v.12 no.1
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    • pp.63-66
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    • 2014
  • The karyotype of F. velutipes Korean cultivar, Fv 3-6, was compared with those of Japanese cultivars, Fv 0-1, Fv 1-5, Fv 11-1, by CHEF gel electrophoresis. The Korean cultivar, Fv 3-6, showed the difference from the three Japanese cultivars in number and size of chromosomes; the Fv 3-6 had two and one more chromosomes then Fv 0-1 and Fv 11-4, and Fv 1-5 had, respectively. The karyotyping by CHEF gel electrophoresis is quite suitable to define new Korean cultivars against Japanese cultivars.

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

  • Park, Sung-Woo;Jin, Kwang-Ho;You, Jae-Hoon;Kim, Dong-Hwan;Seo, Baeseuk;Kim, Young-Sang
    • Analytical Science and Technology
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    • v.11 no.3
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    • pp.213-221
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    • 1998
  • The determination of inorganic anions by capillary zone electrophoresis is reported. A ten component synthetic mixture of anions of bromide, chloride, fluoride, nitrite, nitrate, sulfite, sulfate, perchlorate, chlorate and chlorite was separated by the capillary column and detected by indirect UV method. The running buffer contained 5 mM ammonium dichromate, 10 mM ammonium acetate, 20 mM diethylenetriamine, 10% methanol solution at pH 9.3. A potential of 15 kV at the cathode (reversed polarity) was utilized for the separation of inorganic anions. A complete separation of anions was achieved in less then 10 min and the applicabilities of the method for the analysis of real samples was demonstrated. We compare the concentration of anions in toluene inhaled humen's urine and in postmortem bloods obtained by capillary zone electrophoresis and ion chromatograph.

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

  • Son, Eung-Ryong;Lee, Yong-Se;Yoon, Kyung-Eun;Ha, Yong-Woong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.405-411
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    • 1985
  • The buffer soluble proteins were extracted from six cultivars of barley grains and analyzed by various electrophoresis; 7.5% polyacrylamide slab gel, 2-30% polyacrylamide porosity gradient tube gel, isoelectric focusing (pH4-9) and starch gel electrophoresis. The proteins, esterase, acid phosphatase, malate dehydrogenase, glutamate dehydrogenase and leucine aminopeptidase were investigated to find out the best method to differentiate barley cultivars. The result were that protein and esterase bands in 2-30% polyacrylamide porosity gradient tube gel electrophoresis and protein bands in 7.5% polyacrylamide slab gel electrophoresis showed typical varietal differences. Therefore, those methods were suitable for differentiation of barley cultivars. It was difficult to differentiate the cultivars by the other methodes and patterns of the other enzymes.

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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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    • 2004.08a
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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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    • v.35 no.9
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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.

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

  • 류재천;김현주;서영록;김경란
    • Environmental Mutagens and Carcinogens
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    • v.17 no.2
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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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Analysis of Water-soluble Vitamins in Pharmaceutical Products by Capillary Electrophoresis (모세관 전기영동법을 이용한 제제중의 수용성 비타민 분석)

  • Lee, Jong-Jin;Moon, Dong-Cheul;Lee, Kong-Joo
    • YAKHAK HOEJI
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    • v.38 no.1
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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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Analysis of chromosome aberration, sister chromatid exchange, micronuclei and single cell gel electrophoresis in human lymphocytes exposed in vitro to Bisphenol A and Diethylstilbestrol (비스페놀 A 및 Diethylstilbestrol의 유전독성 평가를 위한 염색체이상, 자매염색분체교환, 소핵형성, 단일세포 겔 전기영동법의 활용)

  • 김병모;정해원
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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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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