• Journal of Ginseng Research
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• 제41권3호
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• pp.326-329
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• 2017

Background: Cultivated ginseng is often introduced as a substitute and adulterant of Russian wild ginseng due to its lower cost or misidentification caused by similarity in appearance with wild ginseng. The aim of this study is to develop a simple and reliable method to differentiate Russian wild ginseng from cultivated ginseng. Methods: The mitochondrial NADH dehydrogenase subunit 7 (nad7) intron 3 regions of Russian wild ginseng and Chinese cultivated ginseng were analyzed. Based on the multiple sequence alignment result, a specific primer for Russian wild ginseng was designed by introducing additional mismatch and allele-specific polymerase chain reaction (PCR) was performed for identification of wild ginseng. Real-time allele-specific PCR with endpoint analysis was used for validation of the developed Russian wild ginseng single nucleotide polymorphism (SNP) marker. Results: An SNP site specific to Russian wild ginseng was exploited by multiple alignments of mitochondrial nad7 intron 3 regions of different ginseng samples. With the SNP-based specific primer, Russian wild ginseng was successfully discriminated from Chinese and Korean cultivated ginseng samples by allele-specific PCR. The reliability and specificity of the SNP marker was validated by checking 20 individuals of Russian wild ginseng samples with real-time allele-specific PCR assay. Conclusion: An effective DNA method for molecular discrimination of Russian wild ginseng from Chinese and Korean cultivated ginseng was developed. The established real-time allele-specific PCR was simple and reliable, and the present method should be a crucial complement of chemical analysis for authentication of Russian wild ginseng.

• 한국건강관리협회지
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• 제4권1호
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• pp.68-74
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• 2006

For identification of four sibling species of the Anopheles hyrcanus complex found in Korea, the 5.8 rDNA-ITS2-28S rDNA region of each species was sequenced and the species-specific primers wee designed The amplified PCR products obtained from each species were analyzed by agarose gel electrophoresis. The result showed a single species- specific band, I.e. 559bp, 432bp, 322bp and 192bp for An. sinensis, An. sp., An. lesteri and An. pullus, respectively. In conclusion, the species-specific PCR primers designed from ITS2 variable regions functioned successfully and specifically, and can be applied as a useful tool for identifying species of the Anopheles hyrcanus complex found in Korea.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권4호
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• pp.212-215
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• 2002

Tyrosinase transcript In the blood Is known as the marker of malignant melanoma and it has been often determined by using reverse transcription-polymerase chain reaction (RT-PCA) . However, after the PCR process, the quantification of amplified CDMA by the gel electrophoresis is not reliable and time-consuming. for this reason, we tried to quantify the PCR product using a cuvette-type biosensor, where the oligonucleotide probe was immobilized on the cuvette surface and the single strand CDMA, the denatured PCH product, was then hybridized onto the immobilized probe to give a response signal. The response was Immediate and takes 15 min to obtain a stable signal. The biosensor was much more sensitive comparing to the gel electrophoresis method. The quantification of PCR product using a cuvette-type biosensor was feasible and rapid.

• International Journal of Industrial Entomology and Biomaterials
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• 제11권2호
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• pp.135-138
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• 2005

Two pairs of DNA primers were designed for the detection of the Zhenjiang (China) strain of Bombyx mori densonucleosis virus (BmDNV-Z). These primers were designed from the nucleotide sequence of major structural protein gene (putative VD1-ORF2). PCR amplification was attempted from different issues (including silk gland, blood, skin and midgut) and feces of the silkworm which infected wit BmDNV-Z were amplified by PCR. Both of the primers gave expected size of in the DNA bands from midgut and feces, but not in the DNA of silk gland, blood and skin. The two bands were sequenced, and their sequence were same as the sequence designed for. BmDNV-Z could be successfully detected in single silkworm after it was infected for 12 hrs, and could not be detected before 9 hrs after infected.

• 대한임상검사과학회지
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• 제39권3호
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• pp.161-166
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• 2007

To develop a rapid and reliable single-tube-based PCR technique for detection simultaneously the quinolone resistance qnrA, qnrB and qnrS genes. After multiple alignment, primers were designed to detect known qnr variants. I was used for A total of 43 extented-spectrum ${\beta}$-lactamases (ESBLs) producing Escherichia coli and Klebsiella pneumoniae isolated from university hospital were tested for screening, as with qnr genes. In optimized conditions, all positive controls confirmed the specificity of the PCR primers. Out of 43 isolates, qnrA genes were detected 19 (44.2%), qnrB genes 5 (11.7%), qnrS genes 15 (34.9%) and 8 (18.6%) isolates were not detected. I report here a fast and reliable technique for rapid screening of qnr positive strains to be used for epidemiological surveys.

• BMB Reports
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• 제29권5호
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• pp.481-483
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• 1996

In competitive PCR, which is used to quantify target DNA an internal standard is needed. Here we present a simple method to construct homologous competitive standards. The method, which is based upon deletion of a portion of the target DNA, does not require any additional primers. This is the simplest method developed thus far to construct a competitive standard. The whole procedure. from construction of a competitive standard to quantitation by PCR, can be completed within a single day.

• Journal of Microbiology
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• 제39권2호
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• pp.126-132
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• 2001

For rapid and secure differentiation of P. infestans from other Phytophthora species, two fragments obtained from randomly amplified polymorphic DNA (RAPD) profiles were selected as markers. Also, primers for in polymerase chain reaction (PCR) to detect P. infestans specifically were developed by analyzing the sequences of ITSII regions in rDNA of Phytophthora species. The primers, PISP-1 and ITS3 amplified a single. Fragment 450 bp of about in P. infestans, but not in other fungal or bacterial isolates. Annealing temperatures and template DNA quantities were varied for the optimization of PCR conditions. From the result of the PCR detection study, species-specific primers were selected under annealing temperatures ranging from 55$^{\circ}C$ to 61$^{\circ}C$, and template DNA levels ranging from 10 pg to 100 ng.

• BMB Reports
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• 제40권2호
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• pp.172-179
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• 2007

PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1928-1935
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• 2015

Microbial growth kinetics is often used to optimize environmental processes owing to its relation to the breakdown of substrate (contaminants). However, the quantification of bacterial populations in the environment is difficult owing to the challenges of monitoring a specific bacterial population within a diverse microbial community. Conventional methods are unable to detect and quantify the growth of individual strains separately in the mixed culture reactor. This work describes a novel quantitative PCR (qPCR)-based genomic approach to quantify each species in mixed culture and interpret its growth kinetics in the mixed system. Batch experiments were performed for both single and dual cultures of Pseudomonas putida and Escherichia coli K12 to obtain Monod kinetic parameters (μ_max and K_s). The growth curves and kinetics obtained by conventional methods (i.e., dry weight measurement and absorbance reading) were compared with that obtained by qPCR assay. We anticipate that the adoption of this qPCR-based genomic assay can contribute significantly to traditional microbial kinetics, modeling practice, and the operation of bioreactors, where handling of complex mixed cultures is required.

• 한국동물위생학회지
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• 제24권4호
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• pp.335-341
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• 2001

Swine dysentery caused by Brachyspira hyodysenteriae, an anaerobic, beta-hemolytic spirochete, is a severe mucohemorrhahic diarrheal disease that primarily affects pigs during the growing and finishing period. The current standard laboratory procedure to culture and identify B hyodysenteriae takes 3 to 7 days. This report present a rapid PCR for detection B hyodysenteriae in a single reaction using DNA from swine intestinal samples. The PCR produced a specific 421bp PCR product with template DNA purified from B hyodysenteriae, and the accuracy for detection of B hyodysenteriae by PCR results compared with those of conventional method was 100% in intestinal specimens. Nonspecific bands were not detected with B innocens, a nonpathogenic common inhabitant spirochete, including other enteric bacterial organisms. This procedure could detect as little as 50 pg of template DNA for B hyodysenteriae.