• Fisheries and Aquatic Sciences
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• v.25 no.5
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• pp.264-275
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• 2022

A non-destructive environmental DNA protocol for the genetic analysis of sea cucumber (Apostichopus japonicus) resources DNA was established. Among the several commercial DNA extraction kits, the DNeasy^® Plant Mini Kit was selected as the best choice to obtain the high-quality genomic DNAs from the mucous sea cucumber. As the temperature and incubation time increased, the amount of extracted environmental DNA was also large, but it was judged that the increased amount did not affect as much as 2-3 times. Therefore, these conditions were not considered to be the main factors to consider in actual environmental DNA extraction. However, the amount of seawater relative to the size of the sample was judged as a major consideration, and a sufficient amount of environmental DNA for analysis was secured when stored within 1 min while stirring the volume of seawater corresponding to the total sea cucumber weight (g). In securing the environmental DNA of sea cucumbers, the mortality rate of sea cucumbers in all experiments was 0, and it was judged that the effects of sea cucumbers were not significant through this treatment. Through the results of this study, sea cucumber DNA research, which has been conducted in a destructive method, can be conducted non-destructively through environmental DNA analysis. Through this study, we have secured a standard protocol that can successfully extract the sea cucumber DNA through environmental DNA. It is not only excellent in terms of time and cost of traditional DNA analysis method currently used, but it is completely non-destructive in the ecosystem of the survey area. It is believed that the system can be transformed in a way that does not affect it. However, it is thought that various standard protocols should be established considering the characteristics of each type.

• Environmental Analysis Health and Toxicology
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• v.29
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• pp.7.1-7.8
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• 2014

Objectives The present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization. Methods A series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method. Results Heating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes. Conclusions Among all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.23 no.2
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• pp.69-89
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• 2020

Scientific trust and quantification of traditional species investigation and results that have been used in ecology for decades has always been a problem and concern for ecologists. Global ecologists have proposed DNA-based species investigation studies to find answers to problems. In this study, we reviewed the global trend of research on environmental DNA(eDNA), which is a method for monitoring species by detecting DNA of organisms naturally mixed in environmental samples such as water, soil, and feces. The first eDNA research confirmed the possibility of species investigation at the molecular level, and commercialization of NGS(Next Generation Sequencing) and DNA metabarcoding elicits efficient and quantitative species investigation results, and eDNA research is increasing in the filed of ecology. In this study, mammals and birds were detected using MiMammal universal primers from 23 samples(3 natural reserves; 20 water bowls) out of 4 patches to verify eDNA for urban ecosystems in Suwon, and eDNA was verified by performing camera trapping and field survey. Most terrestrial species were detected through eDNA, and particularly, mice(Mus musculus), and Vinous-throated Parrotbill (Sinosuthora webbiana) were identified only with eDNA, It has been confirmed to be highly effective by investigating techniques for small and internal species. However, due to the lack of resolution of the primer, weasels(Mustela sibirica) and squirrels(Melanochromis auratus) were not detected, and it was confirmed that the traditional investigation method was effective only for a few species, such as Mogera robusta(Mogera robusta). Therefore, it is judged that the effects of species investigation can be maximized only when eDNA is combined with traditional field survey and Camera trapping to complement each other.

• 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.

• Korean Chemical Engineering Research
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• v.48 no.2
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• pp.147-156
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• 2010

There are four key factors for gas-phase biofilters; biocatalysts(microorganisms), packing materials, design/operating techniques, and diagnosis/management techniques. Biofilter performance is significantly affected by microbial community structures as well as loading conditions. The microbial studies on biofilters are mostly performed on basis of culture-dependent methods. Recently, advanced methods have been proposed to characterize the microbial community structure in environmental samples. In this study, the physiological, biochemical and molecular methods for profiling microbial communities are reviewed, and their applicability to biofilters is discussed. Community-level physiological profile is based on the utilization capability of carbon substrate by heterotrophic community in environmental samples. Phospholipid fatty acid analysis method is based on the variability of fatty acids present in cell membranes of different microorganisms. Molecular methods using DNA directly extracted from environmental samples can be divided into "partial community DNA analysis" and "whole community DNA analysis" approaches. The former approaches consist in the analysis of PCR-amplified sequence, the genes of ribosomal operon are the most commonly used sequences. These methods include PCR fragment cloning and genetic fingerprinting such as denaturing gradient gel electrophoresis, terminal-restriction fragment length polymorphism, ribosomal intergenic spacer analysis, and random amplified polymorphic DNA. The whole community DNA analysis methods are total genomic cross-DNA hybridization, thermal denaturation and reassociation of whole extracted DNA and extracted whole DNA fractionation using density gradient.

• The Plant Pathology Journal
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• v.26 no.3
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• pp.289-292
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• 2010

This study used a modified cetyltrimethylammonium bromide (CTAB) method to efficiently extract DNA from the plant pathogenic fungus Cercospora sojina. Total DNA yield obtained by this method was approximately 1 mg/g of mycelia (fresh weight), and the mean ratio of A260/A280 and A260/A230 were 2.04 and 2.1, respectively. The results of random amplified polymorphic DNA (RAPD) analysis, digestion with restriction enzymes, and Southern hybridization indicated that polysaccharides were effectively removed by this method, and the resulting DNA was sufficient for use in subsequent molecular analysis.

• Journal of the Korean Society of Environmental Restoration Technology
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• v.24 no.6
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• pp.133-144
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• 2021

This study aims to estimate location of land mammals habitat by analyzing spatial data and investigate how to apply environmental DNA monitoring methodology to lotic system in Yangpyeong-gun, Gyeonggi-do. Environmental DNA sampling points are selected through spatial analysis with QGIS open source program by overlaying Kernel density of wild boar(Sus scrofa), elevation, slope and land-cover map, and 81 samples are collected. After 240 mL of water was filtered in each sample, metabarcoding technique using MiMammal universal primer was applied in order to get a whole list of mammal species whose DNA particles contained in filtered water. 8 and 22 samples showed DNA of wild boar and water deer, respectively. DNA of raccoon dog, Eurasian otter, and Siberian weasel are also detected through metabarcoding analysis. This study is valuable that conducted in outdoor lotic system. The study suggests a new wildlife monitoring methodology integrating overlayed geographic data and environmental DNA.

• Environmental Mutagens and Carcinogens
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• v.22 no.2
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• pp.83-89
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• 2002

The single cell gel electrophoresis (comet) assay is one of the useful tools for the study of genetic damage in humans exposed to environmental mutagens and carcinogens. This study was undertaken to evaluate the status of DNA damage in peripheral lymphocytes depending on their sex, age, smoking habits, and other factors in normal healthy Korean population. The 99 volunteers included in the study and out of these, 36 volunteers were smoker and 63 volunteers were non-smoker aged between 20-59 years. All individual answered a questionnaire that assessed their general information including smoking habits and the extent of the environmental tobacco smoke (ETS) exposure, and blood samples were obtained. There was a statistically significant difference in the extent of DNA damage between smoker and non-smoker (p<0.001). A significant difference was also observed between male and female (p<0.001) and amongst the different group of age (p<0.005), however, correlation analysis showed that only smoking habit was a significant factor for DNA damage. No significant effect of smoking duration, number of cigarettes smoking a day, SPY (smoke pack years) in smokers and environmental tobacco smoke exposure in non-smokers on the status of DNA damage was observed.

• Interaction and multiscale mechanics
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• v.4 no.3
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• pp.219-237
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• 2011

Recent advances in scientific computing enable the full atomistic simulation of DNA molecules. However, there exists length and time scale limitations in molecular dynamics (MD) simulation for large DNA molecules. In this work, a two-level homogenization of DNA molecules is proposed. A wavelet projection method is first introduced to form a coarse-grained DNA molecule represented with superatoms. The coarsened MD model offers a simplified molecular structure for the continuum description of DNA molecules. The coarsened DNA molecular structure is then homogenized into a three-dimensional beam with embedded molecular properties. The methods to determine the elasticity constants in the continuum model are also presented. The proposed continuum model is adopted for the study of mechanical behavior of DNA loop.

• Proceedings of the Microbiological Society of Korea Conference
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• 2002.10a
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• pp.22-26
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• 2002

There are a number of ways in which environmental microbiology and microbial ecology will benefit from DNA micro array technology. These include community genome arrays, SSU rDNA arrays, environmental functional gene arrays, population biology arrays, and there are clearly more different applications of microarray technology that can be applied to relevant problems in environmental microbiology. Two types of the applications, bacterial identification chip and functional gene detection chip, will be presented. For the bacterial identification chip, a new approach employing random genome fragments that eliminates the disadvantages of traditional DNA-DNA hybridization is proposed to identify and type bacteria based on genomic DNA-DNA similarity. Bacterial genomes are fragmented randomly, and representative fragments are spotted on a glass slide and then hybridized to test genomes. Resulting hybridization profiles are used in statistical procedures to identify test strains. Second, the direct binding version of microarray with a different array design and hybridization scheme is proposed to quantify target genes in environmental samples. Reference DNA was employed to normalize variations in spot size and hybridization. The approach for designing quantitative microarrays and the inferred equation from this study provide a simple and convenient way to estimate the target gene concentration from the hybridization signal ratio.