• Title/Summary/Keyword: isolating method

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A Simple Method for Extraction of High Molecular Weight DNA fromPorphyra Tenera (Rhodophyta) Using Diatomaceous Earth

  • Kim, Tae-Hoon;Hwang, Mi-Sook;Song, Ju-Dong;Oh, Min-Hyuk;Moon, Yong-Hwan;Chung, Ik-Kyo;Rhew, Tae-Hyoung;Lee, Choon-Hwan
    • ALGAE
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    • v.21 no.2
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    • pp.261-266
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    • 2006
  • The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

Isolation of Deletion Mutants by Reverse Genetics in Caenorhabditis elegans

  • Park, Byung-Jae;Lee, Jin ll;Lee, Jiyeon;Kim, Sunja;Choi, Kyu Yeong;Park, Chul-Seung;Ahn, Joohong
    • Animal cells and systems
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    • v.5 no.1
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    • pp.65-69
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    • 2001
  • Obtaining mutant animals is important for studying the function of a particular gene. A chemical mutagenesis was first carried out to generate mutations in C. elegans. In this study, we used ultraviolet-activated 4,5',8-trimethylpsoralen to induce small deletion mutations. A library of mutagenized worms was prepared for recovery of candidate animals and stored at $15^{\circ}C$ during screening instead of being made into a frozen stock library. In order to isolate deletion mutations in target genes, a polymerase chain reaction (PCR)-based screening method was used. As a result, two independent mutants with deletions of approximately 1.0 kb and 1.3 kb were isolated. This modified and improved reverse genetic approach was proven to be effective and practical for isolating mutant animals to study gene function at the organismal level.

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Advanced Methods for Isolating from and Confirming Campylobacter spp. in Milk and Dairy Products: Review

  • Chon, Jung-Whan;Seo, Kun-Ho;Kim, Binn;Jeong, Dongkwan;Song, Kwang-Young
    • Journal of Dairy Science and Biotechnology
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    • v.38 no.3
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    • pp.121-133
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    • 2020
  • Campylobacter spp. are a type of microaerophilic bacteria that cause human foodborne illnesses worldwide. Among the various types of Campylobacter spp., Campylobacter jejuni and Campylobacter coli account for 90% of foodborne campylobacteriosis. Generally, poultry meats are known to be a primary cause of campylobacteriosis; however, several other types of foods have also been reported to cause campylobacteriosis. Particularly, raw milk has been directly linked to Campylobacter infections among many foodborne illnesses, and cases of campylobacteriosis caused because of the ingestion of unpasteurized raw milk have been recorded worldwide. This review reports (1) general information, history, and nomenclature of Campylobacter spp., (2) epidemiology of Campylobacter spp., (3) detection of Campylobacter spp. from foods including milk and dairy products, and (4) review of methods for controlling the growth Campylobacter spp.

Studies on the Physical Properties of Base Paper for the Manufacture of Mulching Mat for Afforestation Seedling (조림묘목 mulching mat 제조용 base paper의 물리적 특성 연구)

  • Kim, Hyoung-Jin;Oh, Dong-Geun;Yoo, Yeong-Jeong
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.41 no.1
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    • pp.37-43
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    • 2009
  • In this study, the physical properties of base paper for the manufacture of mulching mat for afforestation seedling were investigated. The base paper for mulching mat was prepared by stock conditions of PAE and AKD addition into the screened slurry of KOCC for the strengthening effects of wet tensile and burst strength. The optimum additions of PAE and AKD were considered at 2% and 0.5%, respectively. The accelerated aging by ISO 5630-1 and wet heat aging method under hot water for 2 kinds of commercial mulching and wet strength paper were compared with the base paper prepared for mulching mat manufacture. The accelerated aging test for the base paper prepared for mulching mat manufacture resulted in the same tendency of physical properties as two kinds of commercial products. However, the results of wet heat aging test under hot water indicated that the physical strength for base paper prepared was much higher than others. In addition, the opacity behavior for base paper prepared was enough effects to obstruct weeds growth by isolating transmission of sunlight.

An Application of Electrostatic Repulsion Hydrophilic Interaction Chromatography in Phospho- and Glycoproteome Profiling of Epicardial Adipose Tissue in Obesity Mouse

  • Tran, Trang Huyen;Hwang, In-Jae;Park, Jong-Moon;Kim, Jae-Bum;Lee, Hoo-Keun
    • Mass Spectrometry Letters
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    • v.3 no.2
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    • pp.39-42
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    • 2012
  • Phosphorylation and glycosylation are two of the most important and widespread post-translational modifications (PTMs) in an organism. Proteomics analysis of the PTMs has been challenged by low stoichiometry of the modified proteins and suppression effects by high abundance proteins, typically no-functional house-keeping proteins. In this study, a novel method was applied for not only isolating PTM peptides from intact peptides but also concurrently characterizing of glyco- and phosphoproteome using electrostatic repulsion hydrophilic interaction chromatography (ERLIC) packed with silica coated by crosslinked polyethyleneimine. For 2 mg tryptic digest of mouse proteome of epicardial adipose tissue with fat diet, 802 N-glycosylated peptides of 316 glycoproteins and 159 phosphorylated peptides of 75 phosphoproteins were identified using HPLC chip/quadrupole time-of-flight (Q-OF) tandem mass spectrometer.

Optimal design of Base Isolation System considering uncertain bounded system parameters

  • Roy, Bijan Kumar;Chakraborty, Subrata
    • Structural Engineering and Mechanics
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    • v.46 no.1
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    • pp.19-37
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    • 2013
  • The optimum design of base isolation system considering model parameter uncertainty is usually performed by using the unconditional response of structure obtained by the total probability theory, as the performance index. Though, the probabilistic approach is powerful, it cannot be applied when the maximum possible ranges of variations are known and can be only modelled as uncertain but bounded type. In such cases, the interval analysis method is a viable alternative. The present study focuses on the bounded optimization of base isolation system to mitigate the seismic vibration effect of structures characterized by bounded type system parameters. With this intention in view, the conditional stochastic response quantities are obtained in random vibration framework using the state space formulation. Subsequently, with the aid of matrix perturbation theory using first order Taylor series expansion of dynamic response function and its interval extension, the vibration control problem is transformed to appropriate deterministic optimization problems correspond to a lower bound and upper bound optimum solutions. A lead rubber bearing isolating a multi-storeyed building frame is considered for numerical study to elucidate the proposed bounded optimization procedure and the optimum performance of the isolation system.

Segmentation of Long Chinese Sentences using Comma Classification (쉼표의 자동분류에 따른 중국에 장문분할)

  • Jin Me-Ixun;Kim Mi-Young;Lee Jong-Hyeok
    • Journal of KIISE:Software and Applications
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    • v.33 no.5
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    • pp.470-480
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    • 2006
  • The longer the input sentences, the worse the parsing results. To improve the parsing performance, many methods about long sentence segmentation have been reserarched. As an isolating language, Chinese sentence has fewer cues for sentence segmentation. However, the average frequency of comma usage in Chinese is higher than that of other languages. The syntactic information that the comma conveys can play an important role in long sentence segmentation of Chinese languages. This paper proposes a method for classifying commas in Chinese sentences according to the context where the comma occurs. Then, sentences are segmented using the classification result. The experimental results show that the accuracy of the comma classification reaches 87.1%, and with our segmentation model, the dependency parsing accuracy of our parser is improved by 5.6%.

Development of On-line Performance Diagnostic Program of a Helicopter Turboshaft Engine

  • Kong, Chang-Duk;Koo, Young-Ju;Kho, Seong-Hee;Ryu, Hye-Ok
    • International Journal of Aeronautical and Space Sciences
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    • v.10 no.2
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    • pp.34-42
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    • 2009
  • Gas turbine performance diagnostics is a method for detecting, isolating and quantifying faults in gas turbine gas path components. On-line precise fault diagnosis can promote greatly reliability and availability of gas turbine in real time operation. This work proposes a GUI-type on-line diagnostic program using SIMULINK and Fuzzy-Neuro algorithms for a helicopter turboshaft engine. During development of the diagnostic program, a look-up table type base performance module are used for reducing computer calculating time and a signal generation module for simulating real time performance data. This program is composed of the on-line condition monitoring program to monitor on-line measuring performance condition, the fuzzy inference system to isolate the faults from measuring data and the neural network to quantify the isolated faults. Evaluation of the proposed on-line diagnostic program is performed through application to the helicopter engine health monitoring.

Application of digital polymerase chain reaction technology for noninvasive prenatal test

  • Lee, Seung Yong;Hwang, Seung Yong
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.72-78
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    • 2015
  • Recently, noninvasive prenatal test (NIPT) has been adopted as a primary screening tool for fetal chromosomal aneuploidy. The principle of NIPT lies in isolating the fetal fraction of cell-free DNA in maternal plasma and analyzing it with bioinformatic tools to measure the amount of gene from the target chromosome, such as chromosomes 21, 18, and 13. NIPT will contribute to decreasing the need for unnecessary invasive procedures, including amniocentesis and chorionic villi sampling, for confirming fetal aneuploidy because of its higher positive predictive value than that of the conventional prenatal screening method. However, its greater cost than that of the current antenatal screening protocol may be an obstacle to the adoption of this innovative technique in clinical practice. Digital polymerase chain reaction (dPCR) is a novel approach for detecting and quantifying nucleic acid. dPCR provides real-time diagnostic advantages with higher sensitivity, accuracy, and absolute quantification than conventional quantitative PCR. Since the groundbreaking discovery that fetal cell-free nucleic acid exists in maternal plasma was reported, dPCR has been used for the quantification of fetal DNA and for screening for fetal aneuploidy. It has been suggested that dPCR will decrease the cost by targeting specific sequences in the target chromosome, and dPCR-based noninvasive testing will facilitate progress toward the implementation of a noninvasive approach for screening for trisomy 21, 18, and 13. In this review, we highlight the principle of dPCR and discuss its future implications in clinical practice.

Application of an Efficient Method for Isolation of Mitochondria from Biological Samples

  • Jun, Jin Hyun;Kim, Jihyun;Kim, Kyung Tae;Sung, Ho Joong
    • Biomedical Science Letters
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    • v.20 no.3
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    • pp.180-184
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    • 2014
  • Mitochondria play a crucial role in many essential biological events by way of the electron transport chains and intermembrane proteins that they contain. Abnormalities in the mitochondria are strongly correlated with the development of diseases such as atherosclerosis, cancer, and diabetes. However, the study of mitochondria has been referred to as 'labor-intensive' because of the difficulty in isolating the organelles from their various sources, which can include cultured cells and tissues. Multiple companies provide mitochondria isolation kits, and it is possible for investigators to use different kits and apply different protocols depending on the source of the mitochondria. Therefore, we focused on producing an isolation buffer that could be applied to both cultured cells and tissues, and optimized an isolation protocol that could be used with both. Specifically, we adjusted the buffer condition that can be applied to human cervical cancer cells, fibroblasts, and tissues such as mouse liver and spleen. We also optimized the protocol to improve the efficacy and efficiency of the steps involved in the isolation of mitochondria. These methodological improvements may contribute to advanced research by allowing investigators to overcome the difficulties involved in isolation of mitochondria from biological samples.