• Title, Summary, Keyword: Agarose Standard

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Estimation of Fluid Saturations Using Agarose Standard in NMR Imaging (자기 공명 영상법에서 Agarose 표준 물질을 사용한 유체 포화도의 계산)

  • Kim, Kyung-Hoe
    • Applied Chemistry for Engineering
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    • v.10 no.1
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    • pp.160-165
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    • 1999
  • Agarose gels can be used as reference standards for the measurement of fluid properties in porous media because the relaxation properties of the gel reference standard and those of the fluid in porous media can be closely matched. The use of reference standard to determine porosity and saturation is discussed and the requirements for gel NMR properties given. The relaxtion times of agarose gels measured at 2.0 Tesla are illustrated as a function of agarose and paramagnetic impurity ($CuSO_4$) concentrations. This work shows an empirical result between agarose gel composition and gel relaxtion times. The average value for the porosity distribution is 17.7%, which compares well with the value calculated with the gravimetric analysis. Finally, two phase immiscible displacement using agarose gels as a reference standard was performed. The saturation profiles appear to be consistent with what one might calculate for a one-dimensional displacement in a uniform porous media.

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Comparative Study of the Standard Plaque Assay with Solid-overlay and Immunofocus Assay for Varicella-zoster Virus Titration (수두바이러스의 정량에 있어서 Solid-overlay Standard Plaque Assay와 Immunofocus Assay의 효용성 비교 연구)

  • Lee, Hwa-Kyung;Jeong, Yong-Seok
    • The Journal of Korean Society of Virology
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    • v.30 no.1
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    • pp.61-70
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    • 2000
  • Standard plaque assay using agarose-overlay has long been used for titration of many infectious virus particle. Plaque assay for the titration of varicella-zoster virus and its live vaccine requires three intermittent agarose overlay to visualize plaques. Overall procedure of the assay takes at least nine days from virus inoculation and microbe contamination including fungi is frequently accompanied during incubation period. We studied whether an immunofocus assay in conjunction with peroxidase-mediated immunohistochemical reaction may replace the standard plaque assay for the virus titration by comparing the two methods. A linear relationship was observed between number of foci and virus dilution. The number of foci in a given dilution of virus appeared a little higher than counted plaques formed in standard plaque assay. Independent titration results obtained from two assay methods for a given dilution of virus demonstrated a strong correlation ($r^2=0.99$). Foci of virus infected cells as revealed by the enzyme reaction could be counted either 4 days post-infection (p.i.) under low magnification (40X) microscopy, or 6 days p.i. by naked eye observation. Larger size of cell cuture plate, virus adsorption at $35^{\circ}C$, and 10% FBS in diluent appeared to be better conditions for the assay. Immunofocus assay will be an effective and dependable titration method for varicella-zoster virus and its live vaccine in place of the standard plaque assay in respect to accuracy, costs, and experimental convenience.

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Investigation of the Temperature Change and Quantity of Heat Stimulus of the Commercial Indirect Moxibustion (상용 간접구의 연소형태와 열자극량에 대한 연구)

  • Kwon, O-Sang;Lee, Sang-Hun;Cho, Sung-Jin;Choi, Kwang-Ho;Choi, Sun-Mi;Ryu, Yeon-Hee
    • Journal of Acupuncture Research
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    • v.28 no.6
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    • pp.139-147
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    • 2011
  • Objectives : In this research, relatively the characteristic in the combustion according to the brands of the commercial indirect moxibustion is compared and the commercial indirect moxibustion is standardized and this result tries to be provided as necessary basic data. Methods : After adhering to the agarose gel surface in which the thermocouple is inserted, 6 kinds of commercial indirect moxibustion were burnt off and the burning behaviour of the commercial indirect device and heat quantity of stimulus was compared. Results : 1. The form of combustion did not have a difference in 6 kinds of commercial indirect moxibustion combustion. 2. As to the miximum temperature, 'Seoam' and 'Dongbang' was higher than 'Baekryoung' and 'Taeyang'. 3. It was long so that the highest temperature reaching time of 'Seoam' could note in comparison with the other brands. And the highest temperature reaching time of 'Baekryoung' was short to note in comparison with the other brands. 4. As to the quantity of heat stimulus, 'Seoam' was the biggest and 'Baekryoung' was the smallest. 5. The quantity of heat stimulus of 'Dongbang' was the most stable. Conclusions : In this research, relatively the form of combustion of 6 kinds of commercial indirect moxibustion and heat quantity of stimulus were compared. It desires to anticipate the result that it makes the skin.

A Comparison Study of Signal Intensity of Gadolinium Contrast Media on Fast Spin echo and Ultra Short Time Echo Pulse Sequence at 3T MRI-Phantom Study (3T 자기공명영상 Fast Spin Echo (FSE)와 Ultra Short Time Echo (UTE) 펄스 시퀀스에서 가돌리늄 조영제 희석농도와 신호강도 비교 -팬텀 연구)

  • Lee, Suk-Jun;Yu, Seung-Man
    • Journal of radiological science and technology
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    • v.38 no.3
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    • pp.253-259
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    • 2015
  • The information of contrast media concentration on target organ is very important to get reduce the side effect and high contrast imaging. We investigated alternation of signal intensity as a function of the modality of Gd-based contrast media on spin echo and ultra short time echo (UTE) of T1 effective pulse sequence at 3T MRI unit. Gadoxetic acid, which is a MRI T1 contrast medium, was used to manufacture an agarose phantom diluted in various molarities, and sterile water and agarose 2% were used as the buffer solution for the dilution. The gold standard T1 calculation was based on coronal single section imaging of the phantom mid-point with 2D Inversion recovery spine-echo pulse sequence MR imaging for testing of phantom accuracy. The 1-2mmol/L and 7mmol/L was shown the maximum signal intensity on spin echo and UTE respectively. We confirm the difference of contrast media concentration which was shown the maximum signal intensity depending on the T1 effective pulse sequence.

Specific detection of salmonella enteritidis using polymerase chain reaction method (PCR을 이용한 salmonella enteritidis의 특이적 검출)

  • 조미영;여용구;김영섭;이정학;이병동
    • Korean Journal of Veterinary Service
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    • v.23 no.3
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    • pp.227-233
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    • 2000
  • Salmonella enteritidis is the most prevalent etiologic agents of foodborne acute gastroenteritis. Direct isolation and identification of S enteritidis are time consuming work and not so highly sensitive. This study was conducted to develop for the specific detection of S enteritidis using polymerase chain reaction(PCR). PCR primers were selected to amplify a 351-base pair(bp) DNA fragment from the salmonella plasmid virulence A(spv A) gene of S enteritidis. With the primers, 351 bp DNA products were amplified from S enteritidis but not from other B, D, Cl serogroup Salmonella spp. It was sensitive to detect up to 40 pg of template DNA by agarose gel electrophoresis. This PCR assay is very rapid and specific method and less time consuming than the standard bacteriological methods.

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Isolation and Characterization of Marinobacter sp. KS-1, which Produces a Chondroitin Sulfate-like Mucopolysaccharide

  • Lee, Dae-Sung;Kim, Kyung-Suk;Lee, Myung-Suk;Kim, Young-Mog
    • Fisheries and aquatic sciences
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    • v.13 no.3
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    • pp.210-215
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    • 2010
  • In an effort to isolate a bacterium producing chondroitin sulfate (CS), a marine bacterium, KS-1, which produces mucopolysaccharides, was isolated from seawater and identified as Marinobacter sp. based on analyses of its morphological and biochemical traits and 16S rDNA sequence. Agarose-gel electrophoresis showed that the KS-1 strain produces a CS-like mucopolysaccharide. Structural analysis using Fourier transform infrared spectroscopy revealed that the structure of the CS-like mucopolysaccharide produced by Marinobacter sp. KS-1 is similar to that of dermatan sulfate (CS B). However, the molecular mass of the CS-like mucopolysaccharide is higher than that of standard chondroitin sulfates. Considering the above results, we conclude that the Marinobacter sp. KS-1 produces a CS-like mucopolysaccharide that differs somewhat from CS B in molecular mass.

The Use of the Strain Containing Multiple Plasmids as Size Reference Plasmids (분자량측정을 위한 Reference Plasmid 보유균주의 이용)

  • Bang, Sung-Hyuk;Lee, Yoo-Chul;Seol, Sung-Yong;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.267-273
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    • 1987
  • In the analysis of plasmid profile obtained by agarose gel electrophoresis, the strain harvoring multiple reference plasmids of known molecular weight were needed to estimate the size of unknown molecular weight plasmids. Six strains of E. coli isolated from clinical specimens carried multiple plasmids and these strains could be available as a reference plasmids harvoring strain. These E. coli strains showed 4 to 9 plasmids of various size ranging 2.5 to 94.3 megadalton (Mdal). The correlation coefficients of linear regression between the relative mobility and molecular weight were 0.99966 to 1.00000. Among them, E. coli KE327 from throat which contained 7 plasm ids as follows: 79 Mdal, 46 Mdal, 33 Mdal(pKY3027 C), 4.9Mdal, 3.8Mdal(pKY3027 E), 3.5Mdal, and 2.7 Mdal. Relative amount of the pKY3027 C was the smallest(7.09%) and that of the pKY3027 E was the largest (24.24%) among the plasmid fractions of E. coli KE327.

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Evaluation of Genetic Variability in Kenkatha Cattle by Microsatellite Markers

  • Pandey, A.K.;Sharma, Rekha;Singh, Yatender;Prakash, B.;Ahlawat, S.P.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.12
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    • pp.1685-1690
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    • 2006
  • Kenkatha cattle, a draft purpose breed, which can survive in a harsh environment on low quality forage, was explored genetically exploiting FAO-suggested microsatellite markers. The microsatellite genotypes were derived by means of the polymerase chain reaction (PCR) followed by electrophoretic separation in agarose gels. The PCR amplicons were visualized by silver staining. The allelic as well as genotypic frequencies, heterozygosities and gene diversity were estimated using standard techniques. A total of 125 alleles was distinguished by the 21 microsatellite markers investigated. All the microsatellites were highly polymorphic with mean allelic number of 5.95${\pm}$1.9 (ranging from 3-10 per locus). The observed heterozygosity in the population ranged between 0.250 and 0.826 with a mean of 0.540${\pm}$0.171, signifying considerable genetic variation. Bottleneck was examined assuming all three mutation models which showed that the population has not experienced bottleneck in recent past. The population displayed a heterozygote deficit of 21.4%. The study suggests that the breed needs to be conserved by providing purebred animals in the breeding tract.

Real-time Nucleic Acid Sequence Based Amplification (Real-time NASBA) for Detection of Norovirus

  • Lee, In-Soo;Choi, Dong-Hyuk;Lim, Jae-Won;Cho, Yoon-Jung;Jeong, Hye-Sook;Cheon, Doo-Sung;Bang, Hye-Eun;Jin, Hyun-Woo;Choi, Yeon-Im;Park, Sang-Jung;Kim, Sung-hyun;Lee, Hye-Young;Kim, Tae-Ue
    • Biomedical Science Letters
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    • v.17 no.3
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    • pp.191-196
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    • 2011
  • Noroviruses (noroV) are the major cause of nonbacterial gastroenteritis in humans worldwide. Since noroV cannot yet be cultured in vitro and their diagnosis by electron microscopy requires at least $10^6$ viral particles/g of stool a variety of molecular detection techniques represent an important step towards the detection of noroV. In the present study, we have applied real-time nucleic acid sequence-based amplification (real-time NASBA) for simultaneous detection of NoroV genogroup I (GI) and genogroup II (GII) using standard viral RNA. For real-time NASBA assay which can detected noroV GI and GII, a selective region of the genes encoding the capsid protein was used to design primers and genotype-specific molecular beacon probes. The specificity of the real-time NASBA using newly designed primers and probes were confirmed using standard viral RNA of noroV GI and GII. To determine the sensitivity of this assay, serial 10-fold dilutions of standard viral RNA of noroV GI and GII were used for reverse transcription polymerase chain reaction (RT-PCR) and real-time NASBA. The results showed that while agarose gel electrophoresis could detect RT-PCR products with 10 pg of standard viral RNA, the real-time NASBA assay could detect 100 fg of standard viral RNA. These results suggested that the real-time NASBA assay has much higher sensitivity than conventional RT-PCR assay. This assay was expected that might detect the viral RNA in the specimens which could have been false negative by RT-PCR. There were needed to perform real-time NASBA with clinical specimens for evaluating accurate sensitivity and specificity of this assay.

Polyethylenimine Mediated Gene Delivery with Various Liposomal Formulations (폴리에틸렌이민 및 그들의 리포좀이 중재된 Plasmid DNA의 운반)

  • Han, In Sook;Jun, Mi Sook;Lee, Kab Yong
    • Journal of the Korean Chemical Society
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    • v.43 no.2
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    • pp.193-198
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    • 1999
  • The transfection efficiency of plasmid DNA was inspected using multi-cationic polymer, 5, 10, 25 and 50KD polyethylenimine (PEI). The optimal neutralization ratio of PEI/DNA complexes by agarose assay was 1.5-2.0 (nmol/nmol) without much difference in molecular weight of PEI.In vitro transfection assay, most of PEI-mediated plasmid delivery was better compared to the naked DNA. Especially, 25KD PEI at optimal condition gave higher transfection rather than the standard assay of DEAE-dextran or Lipofectin. To enhance the cell targeting delivery, the liposome formulations were introduced using phospholipids. As a result, PC/PE liposomes increased 2-2.5 times of the transfection efficiency of PEI single or PC/PE single delivery, but not the case of 25KD PEI. Moreover, the DOTAP/PE-introduced PEI delivery reduced the transfection of DOTAP/PE single delivery. All these results proved that the PEI can be used not only good transfectants and but also good DNA condensing agents in neutral/anionic liposome for cell targeting delivery.

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