• 한국동물생명공학회지
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• 제34권4호
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• pp.259-266
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• 2019

Emulsion polymerase chain reaction (PCR) is performed on compartmentalized DNA, allowing a large number of PCR reactions to be carried out in parallel. Emulsion PCR has unique advantages in DNA amplification. It can be applied in many molecular biological assays, especially those requiring highly sensitive and specific DNA amplification. This review discusses the principle of emulsion PCR and its applications in biotechnology. Related technologies are also discussed.

• Journal of Dairy Science and Biotechnology
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• 제34권1호
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• pp.43-49
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• 2016

Emulsion PCR (ePCR) has recently gained interest in the areas of food safety and biotechnology owing to its highly specific and sensitive performance in the amplification of target DNA. To facilitate the applications of ePCR to food safety and biotechnology, this paper describes the principles of ePCR and the factors that should be considered in designing ePCR. In addition, current research and applications related to ePCR are discussed.

• BMB Reports
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• 제46권5호
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• pp.270-275
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• 2013

It is essential to analyze rare mutations in many fields of biomedical research. However, the detection of rare mutations is usually failed due to the interference of predominant wild-type DNA surrounded. Herein we describe a sensitive and facile method of detecting rare point mutation on the basis of allele-specific amplification in emulsion PCR. The identification and selective amplification of rare mutation are accomplished in one-pot reaction. The allele-specific primers coupled on magnetic beads allow the exclusive amplification and enrichment of the mutant amplicons. The productive beads bearing mutant amplicons are subsequently stained with the fluorescent dyes. Thus, the rare point mutations with a percentage as low as 0.1%, can be detected by fluorescent analysis. The relative percentages of mutation among different samples can be roughly accessed by counting the fraction of fluorescent positive beads through flow cytometry.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.426.3-427
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• 2002

The aim of this study was to enhance the transfection efficiency of emulsion-mediated gene expression by using chitosan, Conventional DNA/emulsion complexes and precondensed DNA/emulsion complexes were prepared by adding either naked or precondensed plasmids to cationic emulsion. The zeta potential. TEM, and size of transfection complexes were measured. In vitro transfection efficiency for boty complexes was also studied by several methods: flow cytometer, expression analysis by confocal microscope, RT-PCR, and in addition. (omitted)

• BMB Reports
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• 제44권11호
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• pp.705-712
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• 2011

Advances in the fields of proteomics and genomics have necessitated the development of high-throughput screening methods (HTS) for the systematic transformation of large amounts of biological/chemical data into an organized database of knowledge. Microfluidic systems are ideally suited for high-throughput biochemical experimentation since they offer high analytical throughput, consume minute quantities of expensive biological reagents, exhibit superior sensitivity and functionality compared to traditional micro-array techniques and can be integrated within complex experimental work flows. A range of basic biochemical and molecular biological operations have been transferred to chip-based microfluidic formats over the last decade, including gene sequencing, emulsion PCR, immunoassays, electrophoresis, cell-based assays, expression cloning and macromolecule blotting. In this review, we highlight some of the recent advances in the application of microfluidics to biochemistry and molecular biology.

• 한국식품과학회지
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• 제48권5호
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• pp.454-460
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• 2016

본 논문에서는 E. coli O157:H7, S. Typhimurium, S. aureus와 B. cereus에 대한 ddPCR의 검출 효율과 검출한계를 측정하였으며 동시 검출을 위한 multiplex 검출 가능성을 타진하였다. ddPCR은 PCR mix를 포함한 시료를 15,000-20,000개의 droplet으로 분할하여 PCR 하는 방법으로 droplet reader를 이용하여 droplet의 형광 신호를 계수하였다. 식중독 세균의 표적 DNA를 검출하기 위해 2가지 색의 형광 probe (TaqMan)를 제작하였다. ddPCR은 표적 유전자의 형광 신호를 $100fg/{\mu}L$부터 $10ng/{\mu}L$까지의 DNA를 검출 할 수 있었다. 이후에 두 종류의 식중독 세균에서 프라이머 농도를 달리하여 표적 DNA 증폭 크기의 분포가 서로 다르게 구별할 수 있음을 확인하여 multiplex PCR의 가능성이 있음을 알 수 있었다. ddPCR은 비교적 낮은 검출한계를 가지기 때문에 식품에 적은 농도로 존재하는 식중독 세균의 검출에 활용이 가능할 것으로 생각되었다. 또한 식품의 전처리 조건 확립과 반응조건 확립을 통하여 향후 복잡한 matrix effect를 가지는 식품에서 극 미량의 균 검출도 가능할 것으로 생각되었다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.424-435
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• 2023

본 연구 결과 DPPH 라디칼 소거 활성은 갈랑가 뿌리 70% 에탄올 추출물(AG.E) 100 ㎍/mL 농도에서 81.8%이었으며 ABTS⁺ 라디칼 소거 활성은 AG.E 50 ㎍/mL의 낮은 농도에서 L-Ascorbic acid (AA)와 유사한 99.8%로 확인되었다. 항노화 활성을 측정하기 위하여 collagenase와 elastase 저해 활성을 측정한 결과 둘 모두에서 AG.E는 50 ㎍/mL의 낮은 농도부터 epigallocatechin (EGCG)보다 더 높은 저해 효과를 나타내었다. 특히 500 ㎍/mL 농도에서 EGCG 대비 3배 이상의 저해 효과를 보였다. CCD-986sk 세포내에서 AG.E의 항노화 효과를 검증하기 위해 UVB로 자극한 다양한 실험에서도 우수한 항노화 효과가 얻어졌다. RT-PCR을 통한 유전자 발현 분석 실험에서 COL1A mRNA 발현량은 AG.E 20 ㎍/mL 저농도에서 무첨가 대비 2.90배 증가시키는 결과가 얻어져 항노화관련 우수한 기능성 소재로 개발 가능성이 확인되었다. 제형에 소재의 적용 시 생리활성의 경시적 보전성에 대한 기초 연구로서 AG.E 및 AA등을 안정한 W/O type emulsion에 첨가하여 25 ℃ 항온조에 보관하면서 1일차, 30일차, 60일차에 DPPH와 ABTS+ 라디칼소거 활성을 측정한 결과 모두 경시적으로 항산화 효과가 높은수준 유지됨을 확인하였다.

• Journal of Biosystems Engineering
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• 제38권2호
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• pp.121-128
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• 2013

Purpose: Porcine proliferative enteropathy (PPE), caused by the obligate intracellular bacterium Lawsonia intracellularis, is a widely distributed disease throughout the world causing substantial economic loss. In order to diagnose PPE rapidly, the rapid kit was developed and tested. Methods: In this study, a rapid kit was developed to screen the PPE rapidly at the pig farm. Also, occult blood test with fecal occult blood (FOB) kit was done for detecting the blood in pig feces which might be the evident of hemorrhagic PPE. For developing the kit, we tested fecal samples of PPE infected pigs diagnosed by polymerase chain reaction (PCR) method. Results: With the developed rapid kit, Lawsonia intracellularis was detected in high density emulsion of ileum. On the other hand, the test result of detecting Lawsonia in feces showed too high non-specific response. In addition, nevertheless the FOB test result showed that blood evident could be founded in pig feces, the diagnosing result was not fit to PCR test result, which shows blood in pig feces could be from not only hemorrhagic PPE but also many reasons. Conclusions: To deal with the PPE effectively, it will be better for farmers to screen the PPE in earlier stage with easy and rapid diagnosing tool on farm. This study found out that the rapid kit could detect the Lawsonia intracellularis and hemoglobin in pig feces. However, the non-specific response to negative samples of PPE was too high to use at a pig farm. Further research is needed for lowering the non-specific response with the rapid kit.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2001년도 추계학술대회
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• pp.138-145
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• 2001

The hydrophobic spores of Streptomyces sp. AA8321 isolated from the Antarctic coast displayed demulsification ability. The aerial spores demulsified an emulsion of kerosene/$0.2\%$ Triton X-100 (2:1, v/v) to $50\%$ and $95\%$ within 1 min contact at the concentrations of $5.0{\times}10^7$ and $1.0{\times}10^8$ spores/ml, respectively. A cold shock protein (csp) gene was cloned from the hydrophobic spore- producing Streptomyces sp. AA8321 using PCR. It encoded a low molecular protein with 68 amino acids showing very low homology with previously reported csp genes. Only the sequence of the first six amino acids was just the same and yet others were different. RNA blot analysis indicated that the csp gene was induced by cold shock, i.e., transferring from $30^{\circ}C$ to $10^{\circ}C$, and this cold shock response proposed that the isolated gene be a new type of csp gene.

• Asian Pacific Journal of Cancer Prevention
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• 제15권20호
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• pp.8693-8698
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• 2014

Background: Up-regulation of hsp90 gene expression occurs in numerous cancers such as lung cancer. D,L-lactic-co-glycolic acid-poly ethylene glycol-17-dimethylaminoethylamino-17-demethoxy geldanamycin (PLGA-PEG-17DMAG) complexes and free 17-DMAG may inhibit the expression. The purpose of this study was to examine whether nanocapsulating 17DMAG improves the anti cancer effect over free 17DMAG in the A549 lung cancer cell line. Materials and Methods: Cells were grown in RPMI 1640 supplemented with 10% FBS. Capsulation of 17DMAG is conducted through double emulsion, then the amount of loaded drug was calculated. Other properties of this copolymer were characterized by Fourier transform infrared spectroscopy and H nuclear magnetic resonance spectroscopy. Assessment of drug cytotoxicity on the grown of lung cancer cell line was carried out through MTT assay. After treatment, RNA was extracted and cDNA was synthesized. In order to assess the amount of hsp90 gene expression, real-time PCR was performed. Results: In regard to the amount of the drug load, IC50 was significant decreased in nanocapsulated(NC) 17DMAG in comparison with free 17DMAG. This was confirmed through decrease of HSP90 gene expression by real-time PCR. Conclusions: The results demonstrated that PLGA-PEG-17DMAG complexes can be more effective than free 17DMAG in down-regulating of hsp90 expression by enhancing uptake by cells. Therefore, PLGA-PEG could be a superior carrier for this kind of hydrophobic agent.