• Journal of Oral Medicine and Pain
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• v.20 no.2
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• pp.515-528
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• 1995

The human genomic deoxyribonucleic acid(DNA) was extracted from the pulp, dentin of 22 teeth by clelex, phenol methods. Samples of the tooth-derived DNA were amplified by polymerase chain reaction(PCR), electrophosed for sex determination by detection of X-Y homologus amelogenin gene and D1S80 locus detection The following results have been achieved. 1. Chelex and phenol method are effective to sex determination in the pulp and dentin 2. Chelex method is not suitable for detection of D1S80 locus. 3. Concentration and purity of DNA for teeth using chelex method is lower than using phenol method. From the above investigation, chelex method is simple, rapid for sex determination, but it is not suitable for detection of VNTRs.

• Plant Biotechnology Reports
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• v.4 no.1
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• pp.49-52
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• 2010

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires <15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.10
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• pp.1503-1507
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• 2006

Goats' milk adulteration with cows' milk is becoming a big problem. In the past, the urea-polyacrylamide gel electrophoresis assay with different motility of ${\alpha}S1$-casein has been applied for the identification of cows' milk adulteration. The detection sensitivity is 1.0%. The aim of this study was to develop a faster and more sensitive method to detect cows' milk which may be present in adulterated goats' milk and goats' milk powder. The published primer was targeted at highly conserved regions in bovine mitochondrial DNA (a 271 bp amplicon). This amplicon was cloned and sequenced to further confirm bovine specific sequence. The chelex-100 was used to separate bovine somatic cells from goats' milk or goats' milk powder samples. Random sampling of different brands of goats' milk powder and tablets from various regions of Taiwan showed the adulterated rate was 20 out of 80 (25%) in goats' milk powders and 12 out of 24 (50%) in goats' milk tablets. With this system, as low as 0.1% cows' milk or cows' milk powder in goat milk or goat milk powder could be identified. This chelex DNA isolation approach provides a fast, highly reproducible and sensitive method for detecting the adulteration of goats' milk products.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.179-185
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• 2007

In this study, we used the method of guanidin isothiocyanate and boiling with Chelex-100 resin to extract genomic DNA of Ralstonia solanacearum from soil. It is more efficient than general protocols to remove inhibitory compounds in soil and R. solanacearum on. Then, we applied polymerase chain reaction and DNA enzyme-linked immunosorbent assay (ELISA) to identify and detect pathogen. The fliC gene of R. solanacearum was selected for specific detection of pathogen and primer sets were designed. Among the primer sets, two specific and sensitive primer sets, RsolfliC(forward: 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J. $Sch\ddot{o}nfeld$ et al.) and RS_247 (forward: 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3 designed by this study), were designed to perform nested PCR. Nested PCR primer was labeled with biotin for hybridization between nested PCR product and probe to analyze with DNA ELISA.

• Tuberculosis and Respiratory Diseases
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• v.43 no.1
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• pp.30-37
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• 1996

Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.

• Parasites, Hosts and Diseases
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• v.57 no.1
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• pp.27-31
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• 2019

PCR is known to be the most sensitive method for diagnosing Trichomonas vaginalis infections. This study aimed to compare the sensitivity of a PCR assay for trichomoniasis (HY-PCR) developed in Hanyang University with the use of a Seeplex Ace Detection $Kit^{(R)}$, using urine collected from four Korean men with prostatic disease. Overall, HY-PCR was more sensitive than the Seeplex Kit. The use of Chelex 100 is recommended for DNA isolation in order to increase the sensitivity of the PCR test.

• Food Science of Animal Resources
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• v.37 no.4
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• pp.599-605
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• 2017

Korean native honey (KNH) is much more expensive than European honey (EH) in Korea, because KNH is a favored honey which is produced less than EH. Food fraud of KNH has drawn attention of the government office concerned, which is in need of a method to differentiate between KNH and EH which are produced by the Asiatic honeybee, Apis cerana and the European honeybee, Apis mellifera, respectively. A method to discriminate KNH and EH was established by using duplex polymerase chain reaction (PCR) in this study. Immunochromatographic assay (IC) was examined to analyze the duplex PCR product. The DNA sequences of primers for the duplex PCR were determined by comparing cytochrome C oxidase genes of the two honey bee species. Chelex resin method was more efficient in extracting genomic DNA from honey than the other two procedures of commercial kits. The duplex PCR amplifying DNA of 133 bp were more sensitive than that amplifying DNA of 206 bp in detecting EH in the honey mixture of KNH and EH. Agarose gel electrophoresis and IC detected the DNA of 133 bp at the ratios of down to 1% and 5% EH in the honey mixture, respectively and also revealed that several KNH products distributed by internet shopping sites were actually EH. In conclusion, the duplex PCR with subsequent IC could also discriminate between KNH and EH and save time and labor.

• Analytical Science and Technology
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• v.21 no.5
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• pp.364-374
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• 2008

Antarctic Environmental Monitoring Handbook' was published by COMNAP/SCAR in 2000. The standardized method described in this handbook is recommended for monitoring of antarctic environment. High pressure bomb technique in this guide was used to decompose soil samples. In compliance with this guide book, high pressure bomb technique was applied to decompose the antarctic soil sampled at the King Sejong Station. An Isotope Dilution-Inductively Coupled Plasma-Mass Spectrometry (ID-ICP-MS) was applied to determine mass concentrations of Pb, Cu and Zn in the soil. The accuracy in this method was verified by the analysis of certified reference materials (CRM) of NIST 2702 (marine sediment). The analytical results agreed with certified value within the range from 99.5~100.8%. Matrix separation was necessitated for the determination of Cu and Zn by Chelex 100 ion exchange resin. As a result, the average mass concentrations of Pb, Cu and Zn which are suspected to be caused by anthropogenic pollution were 332.9 mg/kg, 95.6 mg/kg and 115.3 mg/kg, respectively. Those for the metals sampled in the soils of the remote regions from the station were 28.1 mg/kg, 101.8 mg/kg and 115.6 mg/kg, respectively.

• Journal of Radiopharmaceuticals and Molecular Probes
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• v.7 no.2
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• pp.119-125
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• 2021

⁸²Sr for ⁸²Rb generator was produced through the irradiation of the proton beam on the ^nat.RbCI target at the target irradiation facility installed at the end of the Rl-dedicated beamline of the 100 MeV proton linear accelerator of KOMAC (Korea Multi-purpose Accelerator Complex). The average current of the proton beam was 1.2 µA for irradiation time of 150 min. For the separation and purification of the ⁸²Sr from ^nat.RbCI irradiated, Chelex-100 resin was used. The activities of ⁸²Sr in the irradiated ^nat.RbCI target solution and after purification were 45.29 µCi and 43.4 µCi, respectively. The separation and purification yield was 95.8%. As an adsorbent to be filled in the generator for ⁸²Sr adsorption hydrous tin oxide was selected. The adsorption yield of ⁸²Sr into the generator adsorbent was > 99 %, and the total amount of ⁸²Sr adsorbed to the generator was 21.6 µCi as of the day of the ⁸²Rb elution experiment. When the elution amount was 22 mL, the maximum⁸²Rb elution yield was 93.3%, and the elution yield increased as the flow rate increased. After the eluted ⁸²Rb was filled in the correction phantom of the small PET for animals, a PET image was taken. The image scan time was set to 5 min, and the phantom PET image was successfully obtained. As results of impurity analysis on eluted ⁸²Rb using ICP-MS, ^nat.Rb stable isotopes that compete in vivo of ⁸²Rb were identified as undetected levels and were determined to be No-Carrier-Added (NCA).