• Title/Summary/Keyword: DNA추출

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The comparative study of two extraction methods for ancient DNA: silica suspension method and ultracentrifugal concentrator method (고대 유전자에 대한 두 종류의 DNA 분리 방법의 비교 연구: 실리카 현탁액 방법 및 초원심분리 농축 방법)

  • Lee, Eun-jung;Maixner, Frank;Zink, Albert
    • Analytical Science and Technology
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    • v.31 no.2
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    • pp.65-70
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    • 2018
  • This study compared two methods for preparing ancient DNA (aDNA) for the construction of successful shotgun libraries that may be applied to massive parallel sequencing. For the comparative analysis, the DNA of prehistoric rib samples from Hungary was extracted using either a manually prepared silica suspension or the Amicon Ultracel-15 10K ultracentrifugal device (Millipore). After the extraction of the same amount of bone powder (about 150 mg) from three samples by each method, the amount of extracted double-stranded DNA and the subsequent degree of construction of the shotgun library were analyzed. The Amicon device method was rapid and easier to perform and resulted in an approximately 11-fold higher DNA recovery than that obtained using the silica suspension. The shotgun library constructed using DNA templates prepared by the Amicon device was more successful than that constructed from templates isolated using the silica suspension. The comparative study of these two aDNA extraction methods showed that the Amicon device has the advantages of saving time, process simplicity, and high efficiency.

Comparative Evaluation on Qualitative PCR using Different Extraction Methods for Nucleic Acids on Soybean and Corn Processed Foods (대두 및 옥수수 가공식품에서 유전자재조합체(GMO)의 정성 PCR분석을 위한 핵산 추출방법별 비교)

  • 김영찬;이철수;황순욱;김성조;이영옥;윤성원;서정화;남용석
    • Journal of Food Hygiene and Safety
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    • v.18 no.1
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    • pp.6-13
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    • 2003
  • Various kinds of genetically modified organisms (GMO) and processed foods have been developed during recent years. Genetically modified organisms can be classified into several groups as their development methods. Generally, GMO has three foreign DNA regions such as gene expression adjustment region(Promoter), termination region (terminator) and structure gene. Detection of these regions can be done particularly by polymerase chain reaction (PCR). PCR-based detection can virtually be performed for any GMO within short of time. The most important prerequisite for the application of PCR-based detection is to decide abstraction method of efficient nucleic acids. Specially, in the case of processed food, because nucleic acids of foodstuffs are damaged by heat treatment (sterilization), pressure and fermentation, DNA must be extracted ken the samples prior to PCR analysis. Although many DNA extraction protocols are available, they have rarely been compared in a comprehensive method. In this study low widely used commercial and non-commercial DNA extraction methods-DNeasy$^{TM}$, Wizard$^{TM}$, CTAB, phenol/chloroform system-were compared with respect to the quality and yield of nucleic acids and insertion genes.nes.

Decrease of Genotoxicity by Red Ginseng Root Extract (II) -Decrease of MMS- induced Genotoxicity by Red Ginseng Root Extract in Cul tared NIH3T3 Cells (홍삼 추출물에 의한 유전독성 감소효과(II) -배양 NIH3T3 세포에서 MMS에 의한 유전독성의 감소에 미치는 홍삼추출물 처리효과)

  • 차재영;유병수
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.24 no.1
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    • pp.87-99
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    • 1998
  • We have studied the effects of red ginseng root extract on the derease of MMS-induced gemotoxicity in cultured NIH3T3 cells. The increase in survival and the recovery from DNA synthesis inhibition in MMS-treated cells as a function of normal medium incubation time was potentiated, at a rate higher than those in UV-irradiated cells, by the presence of the ginseng extract. The extract also increased the MMS-induced excision repair as determined by unscheduled DNA synthesis. The amount of MMS-induced DNA single strand breads that are accumulated by polymerase inhibitors was increased, but as a rate lower rate than in UV-induced strand break, by the presence of the extract. These results suggest that the red ginseng extract increase MMS-induced repair and could be used as a reagent for protectiong alkylating agent-induced genotoxicity and cytotoxicity.

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Evaluation of protective effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes in the single cell gel electrophoresis assay (단세포 겔 전기영동법을 이용한 사람 림프구 DNA 손상에 대한 복숭아씨 추출물의 방사선 방어효과 평가)

  • Kim, Jin-Kyu;Park, Tae-Won;Lee, Chang-Joo;Chai, Young-Gyu
    • Journal of Radiation Protection and Research
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    • v.24 no.2
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    • pp.93-99
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    • 1999
  • The alkaline single-cell gel electrophoresis (SCGE) assay, called the comet assay, has been applied to the detection of DNA damage from a number of chemical and biological factors in vivo and in vitro. The comet assay is a novel method to assess DNA single-strand breaks, alkali-labile sites in individual cells. The effect of peach kernel extracts on radiation-induced DNA damage in human blood lymphocytes was evaluated by the SCGE assay. The lymphocytes, with or without pretreatment of the extracts, were exposed to 0, 0.1, 0.3, 0.5, 1.0 and 2.0 Gy of $^{60}Co$ gamma ray. Significantly increased tail moment, which was a marker of DNA strand breaks in the comet assay, showed an excellent dose-response relationship. The treatment of the peach kernel extracts reduced the DNA damage by 30 % in irradiated groups as compared to that in non-treated control groups. The result indicates that the extracts shows radioprotective effect on lymphocyte DNA when assessed by the comet assay.

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Effect of Dendranthema indicum Extracts on Cell and DNA Damage Induced by Oxidative Stress (국화추출물이 산화적 스트레스에 의해 유발되는 세포와 DNA 손상에 미치는 영향)

  • Park, Young-Mi;Kim, Jee-In;Lee, Chang-Ho;Lim, Jae-Hwan;Seo, Eul-Won
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1698-1704
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    • 2011
  • In this study, we evaluated the protective effects of ethanol extracts from Dendranthema indicum on cell and DNA damages induced by oxidative stress. Antioxidant activities of D. indicum extracts are higher than scavenging activities of DPPH free radical and hydroxyl radical by 92.8% and 73.8%, respectively, and higher than ferrous iron chelating effects by 59.4%. D. indicum extracts showed a protective effect on oxidative cell damage by inhibiting lipid peroxidation by 90.3% in the control group, and inhibiting expression level of p21 protein by 79.6% for the control group. This means D. indicum extracts have a great protective effect against oxidative stress. DNA fragmentation inhibition in D. indicum extracts were 89.6% for the control group, which makes the movement of DNA tail reduced, and phosphorylation of H2AX was 20.2% of the radical experiment group. This means that D. indicum extracts effectively inhibit DNA fragmentation and H2AX phosphorylation. Taken together, we suggest that ethanol extract from D. indicum has a role as a useful chemopreventor against oxidative damage.

Inhibitory Effect of Korean Fermented Soybean (Chungkookjang) Extract and Genistein Against Trp-P-1 Induced Genotoxicity in HepG2 Cells

  • Song, Eun Jeong;Kim, Nam Yee;Heo, Moon Young
    • Journal of Food Hygiene and Safety
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    • v.32 no.3
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    • pp.171-178
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    • 2017
  • This study evaluated the protective effect of Chungkookjang (CKJ) extract, a Korean traditional fermented soybean product made from Bacillus species in rice straw and boiled soybean, and one of its main flavonoids, genistein, against Trp-P-1 induced cytotoxicity and DNA damage in HepG2 cells. CKJ and genistein exhibited protective effect against Trp-P-1 induced cytotoxicity and Trp-P-1 induced DNA single strand breaks. CKJ and genistein inhibited Trp-P-1 induced CYP1A1 and CYP1A2 transcription in HepG2 cells. Our results indicated that CKJ and genistein have the protective effect against Trp-P-1 induced cytotoxicity and DNA damage. Via inhibiting expression of CYP1A1 and CYP1A2. CKJ can be used as a promising functional food material that prevents the genotoxicity induced by carcinogens produced by the heat treatment of foods such as heterocyclic amines (HCAs) that cause genomic instability.

Antioxidative and Antigenotoxic Activity of Extracts from Rhododendron mucromulatum Turcz. Flowers (진달래 꽃 추출물의 항산화 및 항유전독성 활성)

  • Lee, Bo-Bae;Chun, Ji-Hae;Lee, Suck-Hee;Park, Hae-Ryong;Kim, Jung-Mi;Park, Eun-Ju;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.12
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    • pp.1628-1632
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    • 2007
  • The present study describes the preliminary evaluation of antioxidant activities and antigenotoxic effect of Rhododendron mucromulatum Turcz. flowers (RMF). The samples were prepared by extracting RMF with four different solvents (methanol, ethanol, acetone, and water), and antioxidant properties were evaluated by determining total phenolic contents (TPC), DPPH radical scavenging activity (RSA), and reducing power (RP). Water extract showed the highest total phenol content (328.1 mg/g gallic acid equivalents). Acetone extract showed the most potent RSA and RP. The $IC_{50}$ for RSA and RP in the acetone extract were 78 mg/mL and 454 mg/mL, respectively. The 200 mM $H_2O_2$ induced DNA damage, measured by Comet assay, was inhibited with water, methanol and acetone extract in dose dependent manner in human leukocytes. The inhibition rates were 42, 62, and 52% at the concentrations of 50 mg/mL of water, methanol and acetone extracts, respectively. These results suggest that R. mucromulatum Turcz. has significant antioxidative activity and protective effect against oxidative DNA damage.

A DNA Microextractor Using Crossed Field Electrophoresis (교차 전기영동법을 이용한 극소형 DNA 추출기)

  • Yi Soyeon;Seo Kyoung-Sun;Cho Young-Ho
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.28 no.8 s.227
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    • pp.1135-1139
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    • 2004
  • This paper presents a microextractor for the separation of DNA molecules by their sizes. The DNA microextractor immobilizes the DNA molecules of specific size in the micropillar array by adjusting the period of the crossed electric field, thus providing a starting-point independent target DNA extraction method without separation process monitoring. The DNA microextractor has been fabricated by a three-mask micromachining process. The velocity of three different DNA molecules has been measured at the electric field of E=5V/0.8cm in the fabricated DNA microextractor, resulting in the reorientation times of $4.80{\pm}0.44sec,\;7.12{\pm}0.75sec$, and $9.88{\pm}0.30sec$ for ${\lambda}$ DNA, micrococcus DNA, and T4 DNA, respectively. T4 DNA is trapped in the micropillar array when the crossed electric field of 5V/0.8cm is applied alternately at a 10 second time interval. The present DNA microextractor filters the DNA in a specific size range by adjusting the magnitude and/or the period of the crossed electric field applied in the micropillar array.

Sampling and Extraction Method for Environmental DNA (eDNA) in Freshwater Ecosystems (수생태계의 환경유전자(environmental DNA: eDNA) 채집 및 추출기술)

  • Kim, Keonhee;Ryu, Jeha;Hwang, Soon-jin
    • Korean Journal of Ecology and Environment
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    • v.54 no.3
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    • pp.170-189
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    • 2021
  • Environmental DNA (eDNA) is a genetic material derived from organisms in various environments (water, soil, and air). eDNA has many advantages, such as high sensitivity, short investigation time, investigation safety, and accurate species identification. For this reason, it is used in various fields, such as biological monitoring and searching for harmful and endangered organisms. To collect eDNA from a freshwater ecosystem, it is necessary to consider the target organism and gene and a wide variety of items, such as on-site filtration and eDNA preservation methods. In particular, the method of collecting eDNA from the environment is directly related to the eDNA concentration, and when collecting eDNA using an appropriate collection method, accurate (good quality) analysis results can be obtained. In addition, in preserving and extracting eDNA collected from the freshwater ecosystem, when an accurate method is used, the concentration of eDNA distributed in the field can be accurately analyzed. Therefore, for researchers at the initial stage of eDNA research, the eDNA technology poses a difficult barrier to overcome. Thus, basic knowledge of eDNA surveys is necessary. In this study, we introduced sampling of eDNA and transport of sampled eDNA in aquatic ecosystems and extraction methods for eDNA in the laboratory. In addition, we introduced simpler and more efficient eDNA collection tools. On this basis, we hope that the eDNA technique could be more widely used to study aquatic ecosystems and help researchers who are starting to use the eDNA technique.

Inhibitory Effect of Extract from Acanthocoris sordidus on Oxidative Damage (꽈리허리노린재(Acanthocoris sordidus) 추출물이 산화적 손상에 미치는 억제 효과)

  • Park, Young Mi;Lim, Jae Hwan;Lee, Jong Eun;Seo, Eul Won
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1078-1084
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    • 2014
  • Here, we showed that Acanthocoris sordidus extract inhibited both cell and DNA damage caused by oxidative stress. In a radical scavenging assay, the scavenging activity of the A. sordidus extract against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radicals was 48.9% and 37.8%, respectively, that of ascorbic acid, which was used as a positive control. The ferrous iron chelating activity of the A. sordidus extract was 80.0% compared to that when ethylenediaminetetraacetic acid (EDTA) was used a control. To verify the inhibitory effect of the extract on oxidative cell damage induced by reactive oxygen species (ROS), a lipid peroxidation assay was performed. The results showed that peroxidation was completely inhibited in an extract-treated group compared to a radical-treated group. The level of p21 protein expression was 68.1% that of a control sample. The DNA cleavage-inhibiting property of the A. sordidus extract-treated group was 53.3% that of a control group. Moreover, the phosphorylation of the H2AX protein was reduced to 39.0% of that treated with radical agents, indicating that the extract might inhibit the DNA damage that causes radical oxidation. Taken together, our findings suggest that the A. sordidus extract is effective not only in repressing oxidation by free oxygen radicals and hydroxyl radicals but also in decreasing cell and DNA damage caused by oxidative stress.