• 제목/요약/키워드: DNA preparation

검색결과 153건 처리시간 0.038초

Preparation of Oligonucleotide Arrays with High-Density DNA Deposition and High Hybridization Efficiency

  • Park, Jeong-Won;Jung, Yong-Won;Jung, Young-Hwan;Seo, Jeong-Sun;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • 제25권11호
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    • pp.1667-1670
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    • 2004
  • In DNA microarray produced by DNA-deposition technology, DNA-immobilization and -hybridization yields on a solid support are most important factors for its accuracy and sensitivity. We have developed a dendrimeric support using silylated aldehyde slides and polyamidoamine (PAMAM) dendrimers. An oligonucleotide array was prepared through a crosslinking between the dendrimeric support and an oligonucleotide. Both DNAimmobilization and -hybridization yields on the solid support increased by the modification with the dendrimers. The increase of the immobilization and hybridization efficiency seems to result from a threedimensional arrangement of the attached oligonucleotide. Therefore, our dendrimeric support may provide a simple and efficient solution to the preparation of DNA microarrays with high-density DNA-deposition and high hybridization efficiency.

Encapsulation of Plasmid DNA in Liposomes: Preparation and Characterization

  • Park, Hyo-Min;Lee, Hwa-Jeong
    • 대한약학회:학술대회논문집
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    • 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.1
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    • pp.300.2-300.2
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    • 2003
  • Unlike cationic liposome/DNA complexes, neutralliposomes containing plasmid DNA are stable in blood and does not selectively entrapped in the lung. The objective of this study was to construct neutral liposomes containing plasmid DNA with optimal encapsulation efficiency. Plasmid DNA (pGL2 clone 753,-6 kb) was encapsulated by the freeze/thawing method into liposomes composed of 1-palmitoyl-2-oleyl-sn-glycerol-3-phosphocholine(POPC), didodecyldimethylammonium bromide(DDAB), distaroylphosphatidyl-ethanolamine polyethylene glycol 2000(DSPE-PEG 2000) and DSPE-PEG 2000-maleimide. (omitted)

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Evaluation and modification of alkaline lysis plasmid preparation method from Lactobacillus spp.

  • Lee, Deog-Yong;Seo, Yeon-Soo;Kang, Sang-Gyun;Yoo, Han Sang
    • 대한수의학회지
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    • 제47권2호
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    • pp.157-162
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    • 2007
  • Lactic acid bacteria (LAB) has been regarded as a useful microorganism and tried to manipulate plasmid DNA for increasing the usefulness. Although several methods have been developed to isolate plasmid DNA from Escherichia coli (E. coli), these methods were not sufficient to apply to LAB with exception of O'Sullivan's lysis method. So, we evaluated plasmid DNA extraction from LAB using general E. coli preparation methods and tried to improve the extraction yield and DNA purity by modifying O'Sullivan's alkaline lysis method. To improve the extraction yield, salt and carrier were added to precipitant and those were incubated at $-70{^{\circ}C}$. Only incubation at $-70{^{\circ}C}$ was the effective method of those modifications. Purity of plasmid DNA was improved by two times of each centrifugation and phenol/chloroform extraction. However, DNA was damaged by twice extraction with phenol/chloroform. Also, exclusion of ethidium bromide showed negative effect to purity. Additionally, it was recommended that improvement of the extraction yield may be due to centrifugation at high speed for more time and to dissolving complete DNA pellet before addition of 7.5 M ammonium acetate. Extraction using this modification produced higher quality of plasmid DNA.

Methods for the Extraction of DNA from Water Samples for Polymerase Chain Reaction

  • Jung, Jae-Sung;Lee, Young-Jong
    • Journal of Microbiology
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    • 제35권4호
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    • pp.354-359
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    • 1997
  • Methods for the extraction of DNA from water sample were approximated. Four different procedures of DNA extraction were carried out with pellets obtained from centrifugation of 4 liter water samples. The recovery efficiency and purity of DNA extracted by each method from different sources were compared. DNA yield varied with extraction methods, Method I, which involves enzymatic and freeze-thaw lysis steps and phenol and phenol-chloroform purification of extracted nucleic acid, showed a significantly higher yield and purity than the other methods. The use of glass beads in the DNA extraction methods improved the purity of DNA suitable for PCR. Bovine serum albumin in the PCR reaction mixture was useful in reducing inhibitory effects of contaminants. The efficiency of an extraction method was determined by the detection of the aer of Aeromonas hydrophila with PCR. The lower limit of detection of A. hydrophila from seeded tap water was 2 CFU/ml in PCR when method I was used for DNA preparation.

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DNA 기반 금속 나노 와이어의 제작기술 (DNA Metallization for Nanoelectronics)

  • 한경엽;이정규
    • 공업화학
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    • 제29권3호
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    • pp.253-257
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    • 2018
  • DNA를 기반으로 한 금속 나노와이어는 전기적인 물성은 떨어지지만, 제작 방식이 간단하고, 대면적에서 대량으로 제작할 수 있으며, 유기 반응을 통해 분자 소자 제작의 기판으로도 사용가능한 차세대 재료로 전망된다. 본 총설에서는 DNA 금속화 반응을 이용한 나노와이어의 제작 및 3차원 구조체의 제작 기술에 대해 소개하고, 이와 관련한 연구 현황과 발전 방향에 대해 논의하고자 한다.

Preparation and Atomic Force Microscopy (AFM) Characterization of DNA Scaffolds as a Template for Protein Immobilization

  • Kim, Hyeran;Lee, Hyun Uk;Lee, Jouhahn
    • 한국진공학회:학술대회논문집
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    • 한국진공학회 2014년도 제46회 동계 정기학술대회 초록집
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    • pp.411.2-411.2
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    • 2014
  • The design of DNA nanostructures is of fundamental importance, the intrinsic value of DNA as a building-block material lies in its ability to organize other bio-molecules with nanometer-scale spacing. Here, we report the fabrication of DNA scaffolds with nano-pores (<10 nm size) that formed easily without the use of additives (i.e., avidin, biotin, polyamine, or inorganic materials) into large-scale structures by assembling DNA molecules at near room temperature ($30^{\circ}C$) and low pH (~5.5). Protein immobilization results also confirmed that a fibronectin (FN) proteins/large scale DNA scaffolds/aminopropylytriethoxysilane (APS)/SiO2/Si substrate with high sensitivity formed in a well-defined manner. The DNA scaffolds can be applied for use with DNA-based biochips, biophysics, and cell biology.

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초음파를 이용한 출토 인골 DNA 추출법 연구 (Applied Research of Ultra Sonication for Ancient DNA Preparation of Excavated Human Skeletal Remains)

  • 김윤지;지상현;홍종욱
    • 보존과학연구
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    • 통권29호
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    • pp.137-148
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    • 2008
  • 고고 유적지에서 출토된 뼈에서 추출한 DNA는 부식산과 풀빅산 등의 토양성분과 콜라겐 등 다양한 오염물질이 포함되어 있어 DNA의 추출 및 분석이 매우 어렵다. 본 연구에서는 phenol 추출법, silica 추출법 등 두 가지 대표적인 고대 DNA 추출법의 효율을 DNA 증폭 결과에 의하여 비교하였다. 또한 울트라급의 초음파를 시료 용해에 적용한 후 silica 추출법으로 DNA를 분리한 방법이 기존의 phenol 및 silica 추출법에 비하여 미토콘드리아 DNA와 아밀로제닌 유전자 증폭 결과가 더 우수한 것으로 나타났다.

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Utilization of functionalized magnetic nanoparticles for high throughput DNA separation

  • 장정호
    • 한국재료학회:학술대회논문집
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    • 한국재료학회 2009년도 추계학술발표대회
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    • pp.12.2-12.2
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    • 2009
  • The work describes anoptimized process to highly efficient and convenient preparation in highthroughput magnetic human DNA separation with chemically functionalizedsilica-coated magnetic nanoparticles. The effect of nanoparticle's size and the surface's hydrophilicity change were studied for magnetic DNA separation process, inwhich the optimum efficiency was explored via the function of the amino-groupnumbers, particle size, the amount of the nanoparticles used, and theconcentration of NaCl salt. The DNA adsorption yields were high in terms of theamount of triamino-functionalized nanoparticles used, and the average particlesize was 25 nm. The adsorption efficiency of aminofunctionalized nanoparticleswas the 4-5 times (80-100%) higher compared to silica-coated nanoparticles only(10-20%). DNA desorption efficiency showed an optimum level of over 0.7 M ofthe NaCl concentration. To elucidate the agglomeration of nanoparticles afterelectrostatic interaction, the Guinier plots were calculated from small angleX-ray diffractions in a comparison of the results of electron diffraction TEM,and confocal laser scanning microscopy. Additionally, the direct separation ofhuman genomic DNA was achieved from human saliva and whole blood with highefficiency.

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