• 제목/요약/키워드: DNA preparation

검색결과 153건 처리시간 0.027초

식물의 초경량 조직을 이용한 미토콘드리아의 DNA와 RNA 정제 (Development of a Highly Efficient Isolation Protocol for Mitochondrial DNA and RNA Using Small Scale Plant Tissues)

  • 김경민;임용숙;신동일;설일환
    • 생명과학회지
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    • 제16권2호
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    • pp.240-244
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    • 2006
  • 본 실험에서는 토마토의 종자를 기내 배양하여 얻어진 1g 이하의 무균 잎 조직을 이용하여 미토콘드리아를 분리 정제하여 MitoTracker를 이용하여 세포생물학적으로 확인하였고, 이들의 mt를 이용하여 미토콘드리아 DNA와 RNA를 추출과 검정을 하였다. 또한 고농도의 이온성을 이용하여 미토콘드리아와 mtDNA 및 mtRNA을 추출할 수 있었으며, 식물의 여러 종류에도 사용되어질 수 있을 것이다. mtDNA는 PCR 분석에 의하여 plastid DNA와 혼재되어 있지 않음을 확인하였다. mtRNA는 RT-PCR 분석을 통하여 plastid RNA와 흔재되어 있지 않음을 확인할 수 있었다.

Immunoliposomes Carrying Plasmid DNA : Preparation and Characterization

  • Kim, Na-Hyung;Park, Hyo-Min;Chung, Soo-Yeon;Go, Eun-Jung;Lee , Hwa-Jeong
    • Archives of Pharmacal Research
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    • 제27권12호
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    • pp.1263-1269
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    • 2004
  • The objective of this study was to characterize immunoliposomes carrying plasmid DNA with optimal encapsulation efficiency and antibody density. Plasmid DNA was encapsulated by the freezing/thawing method into liposomes composed of POPC (1-palmitoyl-2-oleoyl-sn-glycerol- 3-phosphocholine), DDAB (didodecyl dimethyl ammonium bromide), DSPE-PEG 2000 (distearoyl phosphatidyl ethanolamine polyethylene glycol 2000) and DSPE-PEG 2000-maleimide. The liposomes carrying plasmid DNA were extruded through two stacked polycarbonate filters, of different pore size, to control the liposome size. Then, rat IgG molecules were conjugated to the liposomes. The immunoliposomes containing plasmid DNA were separated from the free plasmid DNA and unconjugated IgG by Sepharose CL-4B column chromatography. The DNA amount encapsulated was affected by DDAB (cationic lipid) concentration, the initial amount of plasmid DNA between 10 ${\mu}g$ and 200 ${\mu}g$, the total lipid amount and plasmid DNA size, but not significantly by liposome size. By varying the ratio of DSPE-PEG 2000-maleimide to IgG, the number of IgG molecules per liposome was changed significantly.

A Quick and Safe Method for Fungal DNA Extraction

  • Chi, Myoung-Hwan;Park, Sook-Young;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • 제25권1호
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    • pp.108-111
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    • 2009
  • DNA-based studies, including cloning and genotyping, have become routine in fungal research laboratories. However, preparation of high-quality DNA from fungal tissue requires much time and labor and is often a limiting step for high-throughput experiments. We have developed a quick and safe (QS) DNA extraction method for fungi. Time efficiency and safety in the QS method were achieved by using plate-grown mycelia as the starting material, by eliminating phenol-chloroform extraction procedures, and by deploying a simple electric grinder. This QS method is applicable not only to a broad range of microbial eukaryotes, including true fungi and oomycetes, but also to lichens and plants.

Preparation of fluorescent nucleic acids generating unique emission by primer extension reaction using pyrene-labeled deoxyuridine triphosphate derivatives

  • Takada, Tadao;Tanimizu, Yosuke;Nakamura, Mitsunobu;Yamana, Kazushige
    • Rapid Communication in Photoscience
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    • 제3권4호
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    • pp.76-78
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    • 2014
  • Fluorescent nucleic acids were prepared utilizing the polymerase extension (PEX) reaction to incorporate fluorescent molecules. 2'-Deoxyuridine triphosphate (dUTP) derivatives possessing pyrene molecules as fluorophores were synthesized using the aqueous-phase Sonogashira coupling between 5-Iodo-dUTP and acetylene-linked pyrene molecules. The incorporation of the pyrene (Py)-labeled deoxyuridine triphosphates (PyU) into DNA by polymerase was evaluated by polyacrylamide gel electrophoresis, demonstrating that the PyU can work as a good substrate for the PEX reaction. The fluorescent properties of the functionalized DNA prepared by the PEX reaction were characterized by steady-state fluorescence measurements. The Py-conjugated DNA showed typical emission spectra of the pyrene, and the DNA with two pyrene molecules connected to each other by a diethylene glycol linker exhibited a broadened emission attributed to the electronic interaction between the Py molecules.

아미노실란화 철산화물 나노입자를 이용한 Human DNA의 초고속 자성분리 (High Throughput Magnetic Separation for Human DNA by Aminosilanized Iron Oxide Nanoparticles)

  • 강기호;장정호
    • 한국세라믹학회지
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    • 제45권10호
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    • pp.605-609
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    • 2008
  • This work describes the preparation of functionalized magnetic nanoparticles(MNPs) and their bioapplication to human DNA separation. Silica coated MNPs were prepared by changing the volume ratio of tetraethyl orthosilicate(TEOS) for controlled coating thickness on the original nanoparticle of MNPs. The sol-gel process in silica coating on MNPs surface was adapted for relatively mild reaction condition, low-cost, and surfactant-free. And then amino functionalized magnetic nanoparticles were synthesized using amine groups as surface modifiers. The result of adsorption efficiency for human DNA with amino-functionalized silica coated MNPs was calculated as a function of the number of amine groups.

A Simple and Reliable Method for Preparation of Cross-Contamination-Free Plant Genomic DNA for PCR-Based Detection of Transgenes

  • Hwang, Seon-Kap;Kim, Young-Mi
    • BMB Reports
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    • 제33권6호
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    • pp.537-540
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    • 2000
  • A simplified but reliable method was developed for the polymerase chain reaction (PCR)-based detection of genetically modified (GM) plants. The modified CTAB (mCTAB) method enabled us to prepare a high quality of genomic DNA from several hundred plant leaf samples in one day. Using DNA samples prepared from seven dicots and two monocots, approximately 1.75-kb regions spanning 17 S to 25 S ribosomal RNA genes were successfully amplified in a 2X PCR pre-mix containing BLOTTO. Further fidelity assessment of the mCTAB method by PCR analysis with Roundup Ready soybean (RRS) and non-RRS plants showed that the DNA samples prepared alternately from each of two lines were evidently free of cross-contamination. These results demonstrate that the mCTAB method is highly recommended for the rapid detection of transgenes in large numbers of leaf samples from diverse transgenic plants.

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Preparation of Kimchi Containing Bifidobacterim animalis DY-64

  • Chae Myoung-Hee;Jhon Deok-Young
    • Journal of Microbiology and Biotechnology
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    • 제16권3호
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    • pp.431-437
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    • 2006
  • Aero-tolerant microorganisms were isolated from healthy Koreans over the age of 95 years. The microorganisms were then identified based on their morphological and biochemical characteristics and 16S rDNA sequences. The growth properties of the isolated strains were investigated in kimchi. The characteristics of kimchi containing the microorganisms were studied microscopically, physicochemically, and organoleptically. Among 7 aero-tolerant strains, a strain with a 16S rDNA sequence exhibiting 99% homology with Bifidobacterim animalis strain B83 was selected and named B. animalis DY-64. The new strain showed a better acid resistance and salt resistance (p<0.05) than B. animalis ATCC 25527. After 15 days of fermentation in kimchi, the viability of B. animalis DY-64 was about 10%, and the kimchi had a better overall edible quality than conventional kimchi. Thus, it was found that the application of B. animalis DY-64 to kimchi preparation produced a good overall edible quality.