• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.497-513
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• 1995

Human genomic Deoxyribonucleic acid(DNA) was extracted from teeth by boiling, salting-out, phenol, boiling-phenol methods. The author compared DNA concentration and its purity, the accuracy of sex determination and the results of the D1S80 locus detection among above 4 methods. The following results were obtained : 1. DNA concentration was the highest in pulp with salting-out method and DNA purity was higher in pulp with salting-out and phenol methods than other 2 methods. 2. Sex determination was possible using of the pulp and the dentin of the teeth with four methods but, it was impossible in the enamel and some pulp with boiling method. 3. Amplification of D1S80 locus occurred from pulp and dentin with salting-out, phenol, and boiling-phenol methods. 4. There are no differences among the amplification of X-Y homologus amelogenin gene by application of 4 methods and salting-out, phenol methods efficiently makes available to amplification of D1S80 locus. From the investigation DNA extraction, sex determination, amplification of D1S80 locus was successfully accomplished with salting-out, phenol, boiling-phenol methods Therefore above 3 methods are available and applicable as forensic odontology for individual identification.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1166-1172
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• 1998

A method for the isolation and purification of DNA from a red algae, Porphyra was innovated. The innovation of the method consists mainly of three steps that include sodium acetate treatment, chloroform extraction, and 0.2 volume isopropanol precipitation step. The sodium acetate treatment was designed to remove polysaccharide contamination, and the isopropanol step to remove proteins and salts contaminents. Genomic DNA,s of several species(for example, P. tenera, P. yezoensis, P. seriata, and P. pseudolinearis) was successfully isolated by the innovated method. The amount of DNA purified from one g of sample material with the innovated method was 53 g in average. The resulting DNA was characterized to include high molecular weight and showed no nuclease activity. The DNA was pure enough to be digested directly by various restriction enzymes without any difficulties. Porphyra DNA was pure enough and adequate for amplification reaction through the polymerase chain reaction (small nuclear rDNA PCR amplification).

• KSBB Journal
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• v.15 no.4
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• pp.411-413
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• 2000

A method is described for the rapid and simple isolation of genomic DNA from 3 mL culture of Lactobacillus crispatus KLB46 The isolated DNA using this method was shown to be an excellent substrate for restriction endonclease digestion and PCR. The method is expected to be used in gentic manipulation of L. crispatus KLB46.

• 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.

• Journal of the Korean Wood Science and Technology
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• v.46 no.4
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• pp.402-414
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• 2018

Species identification of wood provides important information for archaeology, restoration of cultural assets, preventing illegal logging, and more. Wood species are usually identified based on their anatomical features with the use of a microscope. However, this method may not be able to distinguish between anatomically similar species or subspecies. To overcome this problem, wood species need to be identified at the molecular level using DNA sequencing. However, unlike living plant cells, wood is difficult to pulverize using a mortar, and DNA extraction from dried wood is challenging. To solve these problems, we propose a pretreatment method in which wood is pulverized using 60-grit sandpaper and hydrated with water for 2 days. Using this method, we were able to stably amplify the rpoB gene from the extracted DNA of Pinus rigida. In addition, sequence analysis of the rpoB gene revealed six single nucleotide polymorphisms (SNPs), which classified the rpoB sequences in the genus Pinus into five groups. Our data indicate that although these SNPs were not suitable for species identification, they can potentially be used to determine the origin of different wood subspecies or individual samples of wood.

• Korean Journal of Environmental Biology
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• v.29 no.3
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• pp.138-143
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• 2011

The efficiency of mtCOI amplication after DNA extraction of benthic harpacticoid Tigriopus japonicus s.l. was tested under different conditions depending on fixative (99% Ethanol, or 4% Formalin) and additional chemicals (Ludox or Rose Bengal). Each experimental group by the fixative was subdivided into four groups, respectively: 1) Control (fixative only), 2) processed with Ludox HS40, 3) processed with Rose Bengal, and 4) processed with both Ludox HS40 and Rose Bengal. For the 99% ethanol-fixed sample, overall success rate of amplification by PCR was 96% or above, while for the 4% formalin-fixed one, success rate was much lower than those of ethanol-fixed: 1) Control: 27%, 2) Ludox HS40: 3%, 3) Rose Bengal: 7%, and 4) Ludox HS40 and Rose Bengal: 3%. As a result present study verify that 99% ethanol is a proper fixative for DNA extraction in meiofauna organisms.

• Animal cells and systems
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• v.3 no.1
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• pp.73-78
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• 1999

Although there are several methods for the preparation of mitochondrial DNA (mtDNA) from mammalian tissues, most are relatively long ultracentrifugation or manipulations by a small-scale method. We escribed a rapid method for large-scale extraction of mtDNA from human placental and horse liver tissues. The method is based on the preparation and homogenization of tissues, urification of crude mitochondria by differential centrifugations and isolation of mtDNA by alkaline Iysis. It was improved from Pre-existing methods by replacing some steps with simpler ones and discarding many others. This method gives a high yield of pure mtDNA(approximately 1-5mg from one placenta; ca. 400-600 g wet weight), depending on its sources (fresh tissue gave better results than frozen one). The resulting mtDNA indicated that this method can yield mtDNA in sufficient purity and quantity to identify the direct restriction analysis on agarose gel, random-primed labeling as a probe, and end labeling. Therefore, the method is ideal for obtaining good mtDNA samples to conduct routine restriction fragment length polymorphism (RFLP) analyses of natural populations for genetic studies.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.163-168
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• 1983

The penicillin resistant plasmid DNA was prepared from Streptomyces bobili YS-40, producing penicillinase, by the phenol extraction method and introduced into Bocillus subtilis IAM 12118 by the transformation procedure of Mahler method. The optimal pH and temperature on the transformation was 7.0, 3$0^{\circ}C$ respectively. Above 20 minutes contact of plasmid DNA and recipient cell was shown the high transformation frequency. The transformant of penicillin resistance was proportionally increased as increase of the DNA concentration. The addition of lysine in transformation system increased the transformation frequency about 6-fold and the addition of the chloramphenicol did not affect the transformation frequency.

• Journal of Life Science
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• v.9 no.1
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• pp.15-21
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• 1999

The random amplified ploymorphic DNAs (RAPD) assay is a simple and useful tool in identification of appropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD's have been amplified by the polymerase chain reaction. RAPD of P yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50$\%$) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57$\%$). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.

• Journal of Food Hygiene and Safety
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• v.22 no.4
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• pp.248-256
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• 2007

In this study, the competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Salmonella spp. in pork meat. After comparing three DNA extraction methods, the modified guanidine thiocyanate-phenol-chloroform method was chosen for Salmonella DNA extraction in artificially inoculated pork meat. The previously reported 284-bp invA gene (Rahn et al. Mol. Cell. Probes 1992) was tested for specificity, and 57 Salmonella strains and 24 non-Salmonella strains were evaluated. All Salmonella strains tested were invA positive, and all non-Salmonella strains produced no false positive amplification products. The detection limit achieved was as low as 1,460 colony-forming units (cfu) per 0.1g of pork meat. For cPCR, the invA gene, which features a 82 bp-deletion, was cloned in the pGEM-4Z vector. A known amount of competitor DNA, which has the same primer binding sites, was co-amplified with Salmonella chromosomal DNA from the artificially inoculated pork meat. The cell-number determined by cPCR was approximately equal to the cfu from the most probable number (MPN) method. Finally, the whole procedure took only 5 hr.