• Title/Summary/Keyword: DNA추출

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Comparison of the Efficiency from Raw and Processed Corns by Five Different DNA Extraction Methods (다섯 가지 DNA 추출방법에 의한 옥수수 원료 및 가공시료의 DNA 추출 효율의 비교)

  • Lee, Hun-Hee;Song, Hee-Sung;Kim, Jae-Hwan;Lee, Woo-Young;Lee, Soon-Ho;Park, Sun-Hee;Park, Hye-Kyung;Kim, Hae-Yeong
    • Applied Biological Chemistry
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    • v.48 no.4
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    • pp.331-334
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    • 2005
  • In this study, the effects of five extraction methods for raw and processed corns were compared with respect to the integrity, yields and quality of DNA extracted from them and the results were assessed by PCR analysis. From the comparison of five extraction methods, DNA integrity showed a similar pattern. Amounts of genomic DNA obtained from the five extraction methods varies from $0.25{\mu}g\;to\;234{\mu}g$ per 1 g sample. The DNA yield extracted with CTAB method and DNeasy Plant Maxi kit is greater than that obtained from other extraction methods. These results would be applicable for the selection of an adequate extraction method for specific samples.

Optimized Condition of Genomic DNA Extraction and PCR Methods for GMO Detection in Potato (유전자재조합 감자의 검정을 위한 DNA분리 및 PCR검출의 최적조건 탐색)

  • Shin, Weon-Sun;Kim, Myung-Hee
    • Korean Journal of Food Science and Technology
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    • v.35 no.4
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    • pp.591-597
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    • 2003
  • To compare the quality of genomic DNA extracted from potato for PCR detection, four different methods, such as silica-based membrane method, silica-coated bead method, STE solution treatment, and CTAB-phenol/chloroform method, were evaluated. Also, to remove an excessive carbohydrate from the potato, ${\alpha}$- and ${\beta}$-amylase were used individually and in combination. When used both silica-based membrane method and silica-coated bead method combined with enzymes, the genomic DNAs were extracted from the raw potato with high purity for PCR. However, the silica-coated head method combined with enzyme treatment was the most efficient for extraction of the genomic DNA from the frozen fried potatoes. When applied with STE solution, the highly purified DNA was extracted from the raw potatoes without enzyme treatment in adequate yield for PCR. In cases of processed potatoes, such as frozen-fried potato and fabricated potato chips, CTAB-phenol/chloroform method is mostly feasible for DNA extraction and PCR efficacy at high sensitivity. As the results of PCR amplification, 216bp of PCR product was detected on 2% agarose gel electrophoresis, but any amplicons derived from New leaf and New leaf Y gene was not detected in any sample.

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

  • Kim, Yun-Ji;Jee, Sang-Hyun;Hong, Jong-Ouk
    • 보존과학연구
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    • s.29
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    • pp.137-148
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    • 2008
  • Analyses of ancient DNA (aDNA) from archaeological and historical skeletal material are characterized by low quality. Many soil contaminants such as humic acid, fulvic acid, and bone collagen are often co-extracted with aDNA and inhibit amplification by polymerase chain reaction (PCR). In this study, we compared with two methods of DNA extraction by phenolchloroform extraction and silica-bead extraction. In addition, we applied new protocol, ultra sonication based silica-bead extraction method to extract aDNA from some ancient human skeletal remains. This method was more effective by both mitochondrial DNA (mtDNA) and amelogenin gene amplification.

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Comparison of DNA isolation methods for detection of foodborne pathogens by real-time PCR from foods (식품으로부터 식중독 세균 검출을 위한 Real-time PCR에 적합한 DNA 추출 방법 비교)

  • Koo, Eun-Jeong;Kim, Dongho;Oh, Se-Wook
    • Korean Journal of Food Science and Technology
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    • v.48 no.4
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    • pp.335-340
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    • 2016
  • This study was conducted to find out the most suitable DNA isolation methods for PCR detection of foodborne pathogens. Four DNA isolation methods including Universal Genomic DNA Extraction Kit (TaKaRa), PrepMan Ultra (Applied Biosystems), boiling method and alkaline lysis method (w/PEG) were tested and compared. The Universal Genomic DNA Extraction kit (TaKaRa) was considered as the more efficient isolation method for Escherichia coli O157:H7 and Staphylococcus aureus in lettuce, fish and beef. Meanwhile to detect the foodborne pathogens directly from foods without enrichment, the four different buffers such as double-distilled water, saline, glycine-saline, glycine-saline with Tween-20 and beef extract were also evaluated. As a result, saline was more suitable buffer for E. coli O157:H7. And double-distilled water was more suitable buffer than saline for S. aureus, respectively

Protective Role of Curcuma longa L. Extracts on Hydrogen Peroxide-Induced DNA Damage in Human Leukocytes (산화적 스트레스로 유도된 인체 백혈구 DNA 손상에 대한 울금 추출물의 보호효과)

  • Seo, Bo-Young;Park, Eunju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.5
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    • pp.545-551
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    • 2017
  • Curcuma longa L. (CL) is widely used as a spice and coloring agent in several foods, such as curry and mustard, as well as cosmetics and drugs. In this study, we investigated the protective effects of CL extracted with various solvents [methanol (MC), ethanol (EC), acetone (AC)] on $H_2O_2-induced$ DNA damage in human leukocytes along with total polyphenol contents (TPC) and antioxidant properties. The antioxidant effects of CL were determined by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (RSA) and superoxide dismutase (SOD)-like activity. The preventive effect of CL on oxidative stress-induced DNA damage and DNA repair capacities were assessed using comet assay. MC showed the highest TPC (11.17 g gallic acid equivalents/100 g) and antioxidant properties among the solvent extracts. The $SC_{50}$ for DPPH RSA was MC: 35.0 > AC: 45.8 > EC: $57.8{\mu}g/mL$ and SOD-like activity was MC: 46.6 > EC: 141.5 > AC: $296.4{\mu}g/mL$. In the comet assay, the $ED_{50}$ value of MC showed the highest inhibition ($86.7{\mu}g/mL$) of $H_2O_2-induced$ DNA damage, followed by AC ($110.0{\mu}g/mL$) > EC ($115.8{\mu}g/mL$). Analysis of the percentage of damaged cells showed that repair capacity significantly decreased at 4, 8, and 12 h from $H_2O_2-induced$ oxidative stress in each extract. After 12 h, level of DNA damage recovery was similar to the negative control level. These results suggest that CL has potential antioxidant activity and a protective effect against oxidation-induced DNA damage, and the methanol extract of CL was the most effective.

Inhibitory Effects of Ginger and Garlic Extracts on the DNA Damage (마늘 및 생강추출물의 DNA 손상억제작용)

  • Kang, Jin-Hoon;Ahn, Bang-Weon;Lee, Dong-Ho;Byun, Han-Seok;Kim, Seon-Bong;Park, Yeung-Ho
    • Korean Journal of Food Science and Technology
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    • v.20 no.3
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    • pp.287-292
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    • 1988
  • The inhibition mechanism of DNA damage by lipid peroxidation was studied through the reaction systems of plasmid pBR322 DNA, linoleic acid and the ethanol extracts obtained from ginger and garlic. The DNA damage was greatly inhibited by the addition of ginger and garlic extracts, and their scavenging effects of active oxygens were also great. It is considered that the inhibitory effects of these extracts on the DNA damage are mainly due to their scavenging effects of active oxygen radicals.

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Comparison of methods of DNA extraction from tree nuts (견과류로부터 효율적인 DNA 추출 방법 비교)

  • Suh, Seung-Man;Park, Saet-Byul;Kim, Mi-Ju;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.50 no.4
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    • pp.357-361
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    • 2018
  • This study aimed to explore efficient DNA extraction methods using tree nuts. Four different DNA extraction procedures, including silica membrane method, modified silica method, cetyltrimethylammonium bromide (CTAB) method, and modified CTAB method were examined for their relative efficiency in extracting DNA from pistachio, pine nut, almond, hazelnut, cashew nut, walnut, and peanut. The quality and quantity of the extracted DNA were subsequently assessed by spectrometric measurements, gel electrophoresis, and PCR amplifications. CTAB method was the most appropriate one for extracting DNA from pine nut, cashew nut, pistachio, and peanut. However, it could be replaced by the silica membrane method for walnut and modified CTAB method for almond and hazelnut.

A Review on the Current Methods for Extracting DNA from Soil and Sediment Environmental Samples (토양 및 퇴적토 환경 시료로부터 DNA 추출하는 방법에 대한 고찰)

  • Yoo, Keun-Je;Lee, Jae-Jin;Park, Joon-Hong
    • Journal of Soil and Groundwater Environment
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    • v.14 no.3
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    • pp.57-67
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    • 2009
  • In soil and sediment environment, microorganisms play major roles in biochemical cycles of ecological significant elements. Because of its ecological significance, microbial diversity and community structure information are useful as indexes for assessing the quality of subsurface ecological environment and bioremediation. To achieve more accurate assessment, it is requested to gain sufficient yield and purity of DNA extracted from various soil and sediment samples. Although there have been a large number of basic researches regarding soil and sediment DNA extraction methods, little guideline information is given in literature when choosing optimal DNA extraction methods for various purposes such as environmental ecology impact assessment and bioremediation capability evaluation. In this study, we performed a thorough literature review to compare the characteristics of the current DNA extraction methods from soil and sediment samples, and discussed about considerations when selecting and applying DNA extraction methods for environmental impact assessment and bioremediation capability evaluation. This review suggested that one approach is not enough to gain the suitable quantity and yield of DNA for assessing microbial diversity, community structure and population dynamics, and that a careful attention has to be paid for selecting an optimal method for individual environmental purpose.

Effect of Extracts from Root Bark of Ulmus parvifolia on Inhibition of Growth and DNA Synthesis of Human Cancer Cells. (느릅나무 근피 추출물에 의한 인체 암세포 증식 및 DNA 합성 억제효과)

  • Lim, Sun-Young
    • Journal of Life Science
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    • v.17 no.9 s.89
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    • pp.1232-1236
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    • 2007
  • Growth and DNA synthesis inhibitory effects of extracts from root bark of Ulmus parvifolia on MG-63 human osteosarcoma cells, HT-29 human colon cancer cells and K-562 leukemia cancer cells were studied. The root bark extract of Ulmus parvifolia was extracted with methanol, hot water and juice. The methanol extract showed the highest inhibitory effect on growth of MG-63, HT-29 and K-562 cancer cells by >85%. The treatment of hot water and juice extracts from root bark of Ulmus parvifolia also inhibited growth of the above cancer cells with increasing concentration. DNA synthesis of MG-63 and HT-29 cancer cells was significantly inhibited by adding methanol, hot water and juice extracts from root bark of Ulmus parvifolia with increasing concentration, showing that the inhibitory effect of growth was more effective on HT-29 cancer cells. These results suggest that the methanol extract from root bark of Ulmus parvifolia may have specific active com-pounds on anticancer effect. The hot water extract also showed a strong inhibitory effect on growth of cancer cells, indicating that the active compounds may be stable to heat.

Antioxidant Activities and Inhibitory Effect on Oxidative DNA Damage of Nelumbinis Semen Extracts (연자육 추출물의 항산화 및 산화적 DNA 손상억제 활성)

  • Park, Jae-Ho;Lee, Byung-Gu;Byun, Gwang-In;Kim, Do-Wan
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.55-59
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    • 2010
  • Objective : This study was conducted to investigate the antioxidant activity and inhibitory effect on oxidative DNA damage of Nelumbinis Semen Extracts Methods : Nelumbins semen were extracted with hot-water and ethylacetate (EtOAC). The 1,1-diphenyl-2-picrylhydrazyl (DPPH) and hydroxyl radical scavenging assay and $Fe^{2+}$ chelating assay were performed for antioxidative effect and ${\phi}X$-174 RF I DNA cleavage assay and intracellular DNA damage assay were used for inhibitory effect on intracellular DNA damage. Results : In DPPH, Hydroxyl radical scavenging activity and $Fe^{2+}$ chelating activity of EtOAC extracts were 96.22%, 53.53%, 64.72%, while those of hot-water extracts were 20.86%, 10.72%, 29.74% at $200{\mu}g/m{\ell}$, respectively. In ${\phi}X$-174 RF I plasmid DNA cleavage assay, the protective effects of EtOAC and hot-water extracts against oxidative DNA damage were 76% and 6% at $200{\mu}g/m{\ell}$, respectively. Conclusion : These results indicated that the seed extracts of Nelumbo nucifera can be used as a natural antioxidants, which effectively inhibits the oxidative DNA damage.