• Title/Summary/Keyword: Extraction buffer

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Improved RNA extraction for fruit tree viruses in RT-PCR assay

  • Lee, Sin-Ho;Kim, Hyun-Ran;Kim, Jae-Hyun;Kim, Jeong-Soo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.139.1-139
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    • 2003
  • Tissues from woody plant contain higher amount of phenolic compounds and polysaccharides, which give inhibitory effects on reverse transcriptase and/or Taq ploymerase. The common multiple-step protocols using several additives to inhibit polyphenoic compounds during nucleic acid extraction are time consuming and laborious. Sodium sulfite (Na$_2$SO$_3$) was used as inhibitor of polyphenolic oxidases in extraction buffer and compare it's effect between commercial RNA extraction kit and small-scale double-stranded RNA (dsRNA) extraction by RT-PCR. During nucleic acid extraction procedure, addition of 0.5%-1.5% (w/v) sodium sulfite to Iysis buffer or STE buffer resulted in lighter color change than extracts without sodium sulfite and improve the RT-PCR detection. When commercial RNA extraction kit used, optimal concentration of sodium sulfite were variable according to the host plant. However, using dsRNA as RT-PCR template, 1.5% sodium sulfite in STE buffer improves the detection of both viruses and unspecific amplifications were reduced significantly, Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

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AUTOMATIC TEXTURE EXTRACTION FROM AERIAL PHOTOGRAPHS USING THE ZI-BUFFER

  • Han, Dong-Yeob;Kim, Yong-Il;Yu, Ki-Yun;Lee, Hyo-Seong;Park, Byoung-Uk
    • Proceedings of the KSRS Conference
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    • 2007.10a
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    • pp.584-586
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    • 2007
  • 3D virtual modeling such as creation of a cyber city or landscape, or making a 3D GIS requires realistic textures. Automatic texture extraction using close range images is not yet efficient or easy in terms of data acquisition and processing. In this paper, common problems associated with automatic texture extraction from aerial photographs are explored. The ZI-buffer, which has depth and facet ID fields, is proposed to remove hidden pixels. The ZI-buffer algorithm reduces memory burden and identifies visible facets. The correct spatial resolution for facet gridding is tested. Error pixels in the visibility map were removed by filtering.

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pH Variation In Soils Considering Buffer Capacity during Electrokinetic Extraction (Electrokinetic정화시 토질의 완충능을 고려한 시료내의 pH변화)

  • 오승록;한상재;김수삼;조성호
    • Proceedings of the Korean Geotechical Society Conference
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    • 2001.03a
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    • pp.585-590
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    • 2001
  • Physicochemical phenomena in soils are dependent upon pH when using electrokinetic extraction for the contaminants removal especially for heavy metals. pH variation in soils is affected on H$\^$+/ and OH ̄ ions produced by electrolysis reaction and buffer capacity of soil. High amount of heavy metals are retained in the soils if the soil buffer capacity remains high enough to resist a change in pH. Therefore, accurate pH estimation of soil is important in the application of electrokinetic mechanism for decontamination and understanding of subsurface physicochemical characteristics is also required as well as considering buffer capacity for the enhanced methods application. For these, buffer capacity and pH distribution were measured for the four soils, and also compared with modeling results. The results of buffer modeling were good agreement with experimental data. It is showed that four soils were effected by buffer capacity

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Optimization of Protein Extraction for Lichen Thalli

  • Kondratiuk, Anna S.;Savchuk, Oleksiy M.;Hur, Jae-Seoun
    • Mycobiology
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    • v.43 no.2
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    • pp.157-162
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    • 2015
  • Lichen-forming fungal proteins have been seldom searched due to many difficulties in their extraction. Phenols, quinones, proteases, and other components released during cell disruption have been known to be the greatest challenges related to protein extraction from lichens. To overcome these problems and maintain good electrophoretic resolution and high protein concentration, an extraction buffer containing polyvinylpolypyrrolidone, ascorbic acid, Triton X-100, polyethylene glycol, proteinase, and oxidase inhibitors in sodium phosphate buffer was developed. This extraction buffer showed high efficiency for all lichen species tested in the study.

Evaluation of Available Soil Silicon Extracting Procedures for Oriental Melon (참외 시설재배 토양에 대한 유효규산 추출방법 비교)

  • Cho, Hyun-Jong;Choe, Hui-Yeol;Lee, Yong-Woo;Chung, Jong-Bae
    • Korean Journal of Soil Science and Fertilizer
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    • v.37 no.4
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    • pp.251-258
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    • 2004
  • Soil testing for silicon (Si) in the upland soils has not been sufficiently investigated. The objective of this study was to identify a suitable Si extraction method for upland soils of oriental melon (Cucumis melo L.). Thirty-eight surface soil samples and matured leaf samples were collected from plastic film houses in Sungju, Gyeongbuk province. In the laboratory, six different methods were used for extracting Si from the soils. The methods included 0.5 N HCl extraction, 1 N sodium acetate buffer (PH 4.0) extraction, citric acid 1% extraction, water extraction, Tiis buffer pH 7.0 extraction, and extraction after incubation with water for 1 week. The concentration of dissolved Si in soil extracts from all methods was determined colorimetrically. With 1 N sodium acetate buffer extraction, as the available soil Si increased, the concentration ofSi in oriental melon leaf increased until around $14g\;SiO_2\;kg^{-1}$ was reached in the form of a saturation curve. Also, among the methods studied, extraction with 1 N sodium acetate buffer was the only method provided a significant linear correlation with oriental melon leaf Si content in the range of extractable soil Si lower than the level which inducing Si saturation in oriental melon leaf. These results indicate that 1 N sodium acetate buffer extraction procedure is the best soil Si test method for upland soils of oriental melon. This sodium acetate buffer extraction procedure is rapid and quite well acquainted with scientists and farmers, since the method has been used for routine paddy soil testing.

The controversial points for the assessment of soil contamination related to the change of pH of extraction solution in using partial extraction in standard method in Korea (국내 토양오염 공정시험방법의 용출법 사용시 용출액의 pH의 변화가 토양 오염 평가에 미치는 문제점)

  • 오창환;유연희;이평구;이영엽
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2000.11a
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    • pp.294-297
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    • 2000
  • Heavy metals are extracted from Chonju stream sediment, roadside soils and sediments along Honam expressway, soils and tailings from mining area using partial ectraction in Standard Method, partial ectraction method with maintaining 0.1N of extraction solution and acid digestion. In samples having buffer capacity against acid, 0.1N of extraction solution can not be maintained and pH of extraction solution increases up to 8.0 when partial extraction in Standard Method is used. The averages and ranges of (heavy metals extracted using partial extraction in standard method, HPE)/(heavy metals extracted using partial extraction method with maintaining 0.1N of extraction solution, HPEM) values are 0.506 and 0.145~1.126 in Cd, 0.534~ and 0.078~0.928 in Zn, 0.461 and 0.041~1.715 in Mn, 0.359 and 0.011~0.874 in Cu, 0.195 and 0.018~1.785 in Cr, 0.710 and 0.003~3.075 in Pb, and 0.088 and 1.73$\times$10$^{-5}$ ~0.303 in Fe. These data indicate that the difference between HPE and HPEM is big in the order of Fe, Cr, Cu, Mn, Cd, Zn and Pb. It is quite possible that the partial extraction method in Standard Method of soil in Korea is not adequate for an assessment of contamination in area where buffer capacity of soil will be decreased or lost after a long term exposure of soils to environmental damage.

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Effect of Protein Fractionation and Buffer Solubility of Forage Sources on In Vitro Fermentation Characteristics, Degradability and Gas Production (조사료 자원의 단백질 분획 및 Buffer 추출이 In Vitro 발효 성상, 분해율 및 Gas 생성량에 미치는 효과)

  • Jin, Guang Lin;Shinekhuu, Judder;Qin, Wei-Ze;Kim, Jong-Kyu;Ju, Jong-Kwan;Suh, Seong-Won;Song, Man-Kang
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.32 no.1
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    • pp.59-74
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    • 2012
  • Buffer solubility and protein fractionation were evaluated from the hays (timothy, alfalfa and klein) and straws (tall fescue and rice), and $In$ $vitro$ trial was conducted to examine the effect of buffer extraction on fermentation characteristics, degradability and gas ($CO_2$ and $CH_4$) production. Buffer soluble protein (SP) content and A fraction in total protein were highest in alfalfa hay as 61% and 41.77%, respectively while lowest in rice straw (42.8% and 19.78%, respectively). No difference was observed in B1 fraction among forages but B2 fraction was slightly increased in klein hay (12.34%) and tall fescue straw (10.05%) compared with other forages (6.34~8.85%). B3 fraction of tall fescue was highest as 38.49% without difference among other forages while C fraction was highest in rice straw. pH in incubation solution was higher in all forages after extraction than before extraction at 3h (P<0.01) and 6h (P<0.05), and pH from hays of timothy and alfalfa was higher than the other forages at 6h (P<0.05) and 12h (P<0.001). Regardless of extraction, ammonia-N concentration from alfalfa hay was increased at all incubation times and extraction effect was appeared only at 3h incubation time (P<0.01). Total VFA concentration from alfalfa hay was highest up to 24h incubation while those from tall fescue straw and rice straw were lowest. Buffer extraction decreased (P<0.01~P<0.001) the total VFA concentration. Acetic acid proportion was increased (P<0.001) before extraction of forages but no difference was found between forages. Propionic acid($C_3$) proportion was also increased(P<0.001) before extraction in all forages than in straws at 3h, 24h and 48h incubations, and $C_3$ from hays were mostly higher (P<0.05) than from straws. Butyric acid proportion, however, was not affected by extraction at most incubation times. Parameter 'a' regarding to the dry matter (DM) degradation was increase (P<0.001) in all forages before extraction, and was decreased (P<0.05) in tall fescue straw and rice straw compared with hays. Parameter 'b' was also increased (P<0.001) before extraction but no difference was found between forages. Effective degradability of DM (EDDM) was higher (P<0.001) before extraction in most forages except for rice straw. Buffer extraction decreased (P<0.05) all parameters (a, b, and c) regrading to the crude protein (CP) degradation but no difference was found between forages. Effective degradation of CP (EDCP) was lower (P<0.05) in straws than in hays. Parameters 'a' and 'b' regarding to the NDF degradation (P<0.01) and effective degradability of NDF (EDNDF, P<0.001) were also higher in forages before extraction than after extraction but no difference was found between forages. Buffer extraction reduced (P<0.05~P<0.001) $CO_2$ production from all the forages uo to 24h incubation and its production was greater (P<0.05~P<0.01) from hays than straws. Methane ($CH_4$) production was also greater (P<0.01~P<0.001) in all forages at all incubation times, and its production was greater (P<0.05) from hays than from straws at most incubation times. Based on the results of the current study, it can be concluded that buffer solubility and CP fractionation might be closely related with $In$ $vitro$ VFA concentration, degradability and gas ($CO_2$ and $CH_4$) production. Thus, measurement of buffer solubility and protein fractionation of forages might be useful to improve TMR availability in the ruminants.

Improvement of RT-PCR Sensitivity for Fruit Tree Viruses by Small-scale dsRNA Extraction and Sodium Sulfite

  • Lee, Sin-Ho;Kim, Hyun-Ran;Kim, Jae-Hyun;Kim, Jeong-Soo
    • The Plant Pathology Journal
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    • v.20 no.2
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    • pp.142-146
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    • 2004
  • Woody plant tissues contain great amounts of phenolic compounds and polysaccharides. These substances inhibit the activation of reverse transcriptase and/or Taq polymerase in RT-PCR. The commonly used multiple-step protocols using several additives to diminish polyphenolic compounds during nucleic acid extraction are time consuming and laborious. In this study, sodium sulfite was evaluated as an additive for nucleic acid extraction from woody plants and the efficiency of RT-PCR assay of commercial nucleic acid extraction kits and small-scale dsRNA extraction was compared. Sodium sulfite was used as an inhibitor against polyphenolic oxidases and its effects were compared in RNA extraction by commercial extraction kit and small-scale double-stranded RNA (dsRNA) extraction method for RT-PCR. During nucleic acid extraction, addition of 0.5%-1.5%(w/v) of sodium sulfite to lysis buffer or STE buffer resulted in lighter browning by oxidation than extracts without sodium sulfite and improved the RT-PCR detection. When commercial RNA extraction kit was used, optimal concentrations of sodium sulfite were variable according to the tested plant. However, with dsRNA as RT-PCR template, sodium sulfite 1.5% in STE buffer improved the detection efficiency of Apple chlorotic leaf spot virus (ACLSV) and Apple stem grooving virus (ASGV) in fruit trees, and reduced the unspecific amplifications signi-ficantly. Furthermore, when viruses existed at low titers in host plant, small-scale dsRNA extractions were very reliable.

Shadow Extraction of Urban Area using Building Edge Buffer in Quickbird Image (건물 에지 버퍼를 이용한 Quickbird 영상의 도심지 그림자 추출)

  • Yeom, Jun-Ho;Chang, An-Jin;Kim, Yong-Il
    • Journal of the Korean Society of Surveying, Geodesy, Photogrammetry and Cartography
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    • v.30 no.2
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    • pp.163-171
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    • 2012
  • High resolution satellite images have been used for building and road system analysis, landscape analysis, and ecological assessment for several years. However, in high resolution satellite images, shadows are necessarily cast by manmade objects such as buildings and over-pass bridges. This paper develops the shadow extraction procedures in urban area including various land-use classes, and the extracted shadow areas are evaluated by a manually digitized shadow map. For the shadow extraction, the Canny edge operator and the dilation filter are applied to make building edge buffer area. Also, the object-based segmentation was performed using Gram-Schmitt fusion image, and spectral and spatial parameters are calculated from the segmentation results. Finally, we proposed appropriate parameters and extraction rules for the shadow extraction. The accuracy of the shadow extraction results from the various assessment indices is 80% to 90%.

GaN Film Growth Characteristics Comparison in according to the Type of Buffer Layers on PSS (PSS 상 버퍼층 종류에 따른 GaN 박막 성장 특성 비교)

  • Lee, Chang-Min;Kang, Byung Hoon;Kim, Dae-Sik;Byun, Dongjin
    • Korean Journal of Materials Research
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    • v.24 no.12
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    • pp.645-651
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    • 2014
  • GaN is most commonly used to make LED elements. But, due to differences of the thermal expansion coefficient and lattice mismatch with sapphire, dislocations have occurred at about $109{\sim}1010/cm^2$. Generally, a low temperature GaN buffer layer is used between the GaN layer and the sapphire substrate in order to reduce the dislocation density and improve the characteristics of the thin film, and thus to increase the efficiency of the LED. Further, patterned sapphire substrate (PSS) are applied to improve the light extraction efficiency. In this experiment, using an AlN buffer layer on PSS in place of the GaN buffer layer that is used mainly to improve the properties of the GaN film, light extraction efficiency and overall properties of the thin film are improved at the same time. The AlN buffer layer was deposited by using a sputter and the AlN buffer layer thickness was determined to be 25 nm through XRD analysis after growing the GaN film at $1070^{\circ}C$ on the AlN buffer CPSS (C-plane Patterned Sapphire Substrate, AlN buffer 25 nm, 100 nm, 200 nm, 300 nm). The GaN film layer formed by applying a 2 step epitaxial lateral overgrowth (ELOG) process, and by changing temperatures ($1020{\sim}1070^{\circ}C$) and pressures (85~300 Torr). To confirm the surface morphology, we used SEM, AFM, and optical microscopy. To analyze the properties (dislocation density and crystallinity) of a thin film, we used HR-XRD and Cathodoluminescence.