• Title/Summary/Keyword: Extraction buffer

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Convenient Genetic Diagnosis of Virion Captured (VC)/RT-PCR for Rice Viruses (RSV, RBSDV) and Small Brown Plant Hopper (벼 바이러스(RSV, RBSDV)와 애멸구의 간편한 VC/RT-PCR 유전자 진단기술)

  • Kim, Jeong-Soo;Lee, Su-Heon;Choi, Hong-Soo;Cho, Jeom-Deog;Noh, Tai-Whan;Kim, Jin-Young
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.57-62
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    • 2009
  • Genetic diagnosis method of Virion Captured (VC)/RT-PCR for Rice stripe virus (RSV) and Rice black-streaked dwarf virus (RBSDV), Korean major rice viruses transmitted by small brown plant hopper, Laodelphax striatellus, was developed. Virion extraction buffer for rice plant was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite. However, the extraction buffer for L. striatellus was 0.01M potassium phosphate buffer, pH 7.0, containing 0.5% sodium sulfite and 2% polyvinylpyrrolidone wt 40,000 (PVP-40). Specific primers for detection of RSV and RBSDV were selected for VC/RT-PCR method. The specific primers were used as a duplex primer to detect viruliferous small brown plant hopper collected from Gimpo, Pyeongtaek and Siheung areas in Gyeonggi province. The genetic diagnosis methods of single and duplex VC/RT-PCR for RSV and RBSDV could be used easily and economically, especially on the diagnosis of L. striatellus. The rate of viruliferous insect (RVI) for RSV was compared with ELISA and VC/RT-PCR for L. striatellus collected from fields. RVI by ELISA was same as 9.2% with RVI by VC/RT-PCR. However, there were some different detection results between the methods. It could be suggested that there is a possibility of serological and/or genomic differences among RSV isolates. The portion of RVI detected simultaneously by ELISA and VC/RT-PCR was 71.0%, and the detection rate from VC/RT-PCR was higher as 3.2% than that from ELISA, which had a reason of simultaneous detection ability both RSV and RBSDV of VC/RT-PCR.

Improvement of the Detection Technique of Listeria monocytogenes through Modification of the Enrichment Medium and DNA Extraction Buffer (증균배지 및 DNA 추출법 개량을 통한 Listeria monocytogenes의 검출기법 개선 연구)

  • Lee, Jeeyeon;Seo, Yeongeun;Yoon, Yohan
    • Journal of Food Hygiene and Safety
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    • v.35 no.4
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    • pp.334-340
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    • 2020
  • In this study we developed an enrichment medium and lysis buffers to detect Listeria monocytogenes in meat and processed meat products under various lysis conditions. The enrichment efficiency of L. monocytogenes medium listed in the Food Standards was compared, and thus, Listeria Enrichment Broth (LEB) was modified by adding supplements such as carbon source and minerals. The lysis buffers were developed to extract L. monocytogenes DNA quickly and efficiently under various lysis conditions. L. monocytogenes was most rapidly grown in LEB containing 0.1% pyruvate and 0.1% ferric citrate. A lysis buffer mixed with 0.5% or 1% N-lauroylasrcosine sodium salt, 0.5 N NaOH and 0.5 M EDTA for 30 min at room temperature was found to be the best in terms of DNA purity and yield. These results indicate that developed enrichment medium and lysis buffer can be used to detect L. monocytogenes in meat and processed meat products rapidly and efficiently.

Preparation of Gemcitabine-Loaded PLLA/Gemcitabine-PLLA Microparticles Using Supercritical Fluid and Their Release Characteristics (초임계 유체를 이용한 젬시타빈 함유 PLLA/젬시타빈-PLLA 미립자의 제조 및 방출 특성)

  • Hong, Jin-Hee;Lim, Giobin;Ryu, Jong-Hoon
    • KSBB Journal
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    • v.29 no.6
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    • pp.405-413
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    • 2014
  • In this study, gemcitabine (Gem)-Poly(L-lactic acid) (PLLA) conjugates were synthesized through an amide linkage reaction. Then, the microparticles of Gem-PLLA/PLLA blends containing gemcitabine were prepared using a supercritical fluid process, called aerosol solvent extraction system (ASES). Gemcitabine-loaded Gem-PLLA/PLLA microparticles obtained from the ASES process showed a spherical shape. The amount of gemcitabine released after 30 day incubation in a phosphate buffer solution of pH 7.4 was about 90% of the total amount of gemcitabine present in the product.

Spectrophotometric Determination of Ion Pair Extraction of Quaternary Amines with Metanil Yellow (Metanil Yellow에 의한 4급 Amine류의 Ino Pair 추출 흡광도 정량 (II) - Benzalkonium Chloride, Cetylpyridinium Chloride 및 Dimenhydrinate의 정량)

  • 김영수;최종환
    • YAKHAK HOEJI
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    • v.35 no.1
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    • pp.15-21
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    • 1991
  • Quaternary amines which are widely used as medicines are nitrogen compounds. Metanil yellow(MY) and benzalkonium chloride(BKC) were mixed and added to 10ml of the buffer solution and then the solution was shaken for 5 minutes. The maximum absorption wavelength of the reaction product was 402 nm. Dichloromethane was the best extracting solvent among the several organic solvents and the most suitable pH range was 2~8. When the BKC-MY calibration curve was made in the best experimental condition, the Lambert-Beer's law was obeyed in the range of BKC concentration of 2$\times$$10^{-6}$~9$\times$$10^{-6}$M by UV spectrophotometer. This method was possible to determine quaternary ammonium salts in the pharmaceutical preparation.

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Effect of True Partition Coefficient on the Determination of Extraction Coefficient of Ion-Pair Complexes (이온대 화합물의 참분배계수가 축출정수 결정에 미치는 영향)

  • 심창구
    • YAKHAK HOEJI
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    • v.27 no.2
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    • pp.125-131
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    • 1983
  • Whether ($AB_{w}$) may be neglected against ($A_{w}^{+}$) in the calculation of the extraction coefficient of ion-pairs was criticized by both experiments and theoretical consideration, where ($AB_{w}$) and ($A_{w}^{+}$) mean the molar concentration of ion-pair AB and cation $A^{+}$ in the aqueous phase. Ion-pair complexes were partitioned between phosphate buffer (pH 7.4) and n-octanol. Tetrabutylammonium, isopropamide and methylene blue were selected as cations and benzoic acid, p-toluenesulfonic acid, salicylic acid and taurodeoxycholic acid were selected as counter ions (anions). As a result, conventional methods which assume no existence of ($AB_{w}$) were proven to lack generality. The equation proposed in my earlier report was confirmed to be valid as a general method.

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The Simultaneous Analysis of Oxytetracycline, Chloramphenicol and Sulfamethoxazole in Pork by HPLC (HPLC를 이용한 돈육 중의 Oxytetracycline, Chloramphenicol 및 Sulfamethoxazole의 동시검출)

  • 조혜연;조진국;이치호
    • Food Science of Animal Resources
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    • v.21 no.1
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    • pp.64-70
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    • 2001
  • The extraction procedure and HPLC condition were modified to analyze the residues of oxytetracycline, sulfamethoxazole and chloramphenicol in pork, simultaneously. The antibacterial agents in pork were extracted with 0.02M EDTA-Mcilivine buffer:ethanol:acetonitrile (5:3:2). After the removal of fat with n-hexane, the extracts were evaporated and purified with Sep-pak $C_{18}$ cartridge column using 0.01M oxalic acid 0.1% (v/v) triethylamine (TEA) in acetonitrile. The peak of antibacterial agents was detected with $\mu$ Bondapak C18 column, UV detector (280nm) and 0.01M oxalic acid: methanol: acetonitrile (7.5:2.0:0.5). Detection limits for three antibacterial standards were 0.03 ppm. Calibration curves were linear between 0.03 and 2.0 ppm (R$^2$>0.999). When spiked the level of 1.0 ppm of oxytetracycline, sulfamethoxazole and chloramphenicol into meats, the recoveries from meats were 77.3%, 79.7% and 59.3%, respectively. These results showed that the modified extraction method provided good analytical resolution and the recoveries of the above antibacterial agents in meats.

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A simple method for detection of CMV viral RNAs and satellite RNAs in Korean pepper.

  • J.H. Sung;Park, J.H.;H.Y. Shin;M.U. Chang;H. Sayama;H. Atarashi
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.150.3-151
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    • 2003
  • To analyze the genome of Cucumber mosaic virus(CMV) in pepper, we developed a new extraction method for double-stranded RNA(dsRNA). To isolate the dsRNA, 0.1g of pepper leaves homogenized with 1ml of 5${\times}$EXB extraction buffer[0.5M glycin, 0.5M NaCl, 5mM EDTA(pH9.0/NaOH), 10% Sodium N-lauryl salcosinate(NLS), 10% Sodium dodecylsulfate(SDS)] and purified with the 1/4 volume of phenol: chloroform: isoamylalcohol(25:24:1). dsRNAs from the aqueous phase was precipitated with isopropanol. This procedure was able to detect a minimal amount of dsRNA from CMV infected plant tissue and to distinguish different CMV satellite RNAs by polyacrylamide gel electrophoresis(PAGE). Moreover, this method can be applied CMV infected in pepper or Rice dwarf virus (RDV) infected rice.

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Application of Synergistic Solvent Extraction by Formation of Ternary Complex for Determination of Trace Zn(II) in Water Samples

  • Choi, Jong-Moon;Park, Hyun-Mo;Choi, Sun-Do
    • Bulletin of the Korean Chemical Society
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    • v.27 no.4
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    • pp.563-567
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    • 2006
  • The application of a synergistic solvent extraction by the formation of ternary complex with pyrocatechol violet (PV) and benzalkonium chloride (BC) was studied for determination of trace Zn(II) in water samples. The pH of sample solution and the amount of PV and BC added were optimized for the formation of the stable complex, a proper solvent was selected for the effective extraction, and the concentration of nitric acid was fixed for the back extraction of the complex from the solvent. After the ionic strength of 100 mL sample solution was adjusted to 0.1 M by adding NaCl and the pH was fixed at 9 with a carbonate buffer, 1.0 mL of 2% PV solution was added to form Zn(II)-PV complex then the Zn(II)-PV/BC ternary complex was made by adding 1.0 mL of 10% BC solution. The ternary complex was extracted into 10 mL of MIBK. And the ternary complex was back-extracted with 10 mL of 1.0 mol/L nitric acid to determine Zn(II) by a flame atomic absorption spectrophotometer (flame-AAS). The interference of concomitant ions on the extraction of Zn(II) was investigated. This procedure was applied to the analysis of three real samples such as Dalbang-dam water, laboratory tap water and Jungnajin seawater. The recoveries of Zn(II) in spiked samples were 86.58-104.1%.

Further Purification of Radioprotective Ginseng Protein Fraction by Gel Filtration (Gel filtration에 의한 한방사선 인삼단백 분획의 정제)

  • 김춘미;박경애
    • Journal of Ginseng Research
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    • v.13 no.2
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    • pp.254-259
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    • 1989
  • A radioprotective ginseng protein fraction was obtained from Korean white ginseng powder by the following isolation and purification procedures: Tris-HCI buffer extraction, 70% ammonium sulfate fractionation, CM-rellulosr column chromatography, heat inactivation and Sephadex G-75 column chromatography. This fraction was further purified by Sepharose 4B and Sephadex G-150 column chromatographies. Three fractions obtained were subjected to Native-PAGE and SDS-PAGE using gradient gels and the silver staining method. Molecular weights of the native proteins and their subunits were estimated.

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Moving Object Surveillance System based on Image Subtraction Technique (영상 Subtraction을 이용한 이동 물체 감시 시스템)

  • 이승현;류충상
    • Journal of the Korean Society of Safety
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    • v.12 no.3
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    • pp.60-66
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    • 1997
  • In this paper, a moving object surveillance system, which can extract moving object in real-time, using image subtraction method is described. This technique based on the novelty filter having the structure of neural network associative memory. Digital arithmetic and timing control parts were composed of hardwired controller to treat two-dimensional massive image information. SRAMS having 20 ns access time were used for the image buffer that has high speed write/read property. Image extraction algorithm is discussed and supported by simulation and experiments.

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