• Title/Summary/Keyword: random amplified polymorphic DNA (RAPD-PCR)

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Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis (탄저균의 Random Amplified Polymorphic DNA-PCR 분석)

  • 김성주;박경현;김형태;조기승;김기천;최영길;박승환;이남택;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.56-60
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    • 2001
  • Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.

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Random Amplified Polymorphic DNA (RAPD) Variation in Porhyra yezoensis and P. tenera

  • Beom-Kyu Kim;Gyu-Hwa Chung;Yuji Fujita
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.321-326
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    • 1997
  • The random amplified polymorphic DNA (RAPD) technique was used to anlayze six isolates of two species of Porphyra, P. yezoensis and P. tenera. Four 21-mer prrmers were combined randomly into six groups of double primers and screened for DNA amplification using nuclear and chloroplast tempate DNA. The RAPD patterns resulting from RnRc and CnCc primers provided evidence for both genetically homo-and heterogeneous populations of P. yezoensis and P. tenera. Similarity values obtained by RnRc primer analysis of nuclear DNA varied from 0.364 to 0.714 and those of chloroplast DNA were high, ranging from 0.727 to 1.000, except for P. yezoensis (Enoura).

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Classification of Capsicum annuum Germplasm Using Random Amplified Polymorphic DNA (RAPD를 이용한 고추(Capsicum annuum) 유전자원의 분류)

  • Nam, Seung-Hyun;Choi, Geun-Won;Yoo, Il-Woong
    • Horticultural Science & Technology
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    • v.16 no.4
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    • pp.503-507
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    • 1998
  • This study was initiated to evaluate genetic relationship among various domestic and exotic pepper accessions using random amplified polymorphic DNA(RAPD) markers. The results suggested that the optimum conditions for PCR with random primers in Capsicum spp. could be obtained with 3mM of $MgCl_2$, 1.5U of Taq. DNA polymerase, 10ng of template DNA, $200{\mu}M$ of dNTPs, 200nM of random primer, and $42^{\circ}C$ of annealing temperature. Sixteen random primers showing high band intensity and reproducibility were selected from 80 random primers. Primers having 70% GC content were more effective in DNA amplification than primers having 60% GC content. The total 93 DNA bands including 71 polymorphic bands and 22 monomorphic bands were obtained with selected 16 random primers for 31 pepper cultivars and lines. About 4.4 polymorphic bands per primer were produced. Similarity coefficients were calculated by using 71 polymorphic bands and dendrogram based on the similarity coefficient showed clear classification of 31 peppers into three Capsicum species of Capsicum annuum, Capsicum chinense and Capsicum chacoense.

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Genomic Polymorphisms of Genome DNA by Polymerase Chain Reaction-RAPD Analysis Using Arbitrary Primers in Rainbow Trout (PCR-RAPD 기법에 의한 무지개송어 Genome DNA 의 다형현상)

  • Yoon, J.M.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.303-311
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    • 1999
  • Nuclear DNA was isolated from the sperm cells representing genetic characteristics and genomic polymorphisms of rainbow trout by polymerase chain reaction(PCR) amplification of DNA using arbitrary primers. Genomic DNA fingerprints were generated from rainbow trout sperm DNA by polymerase chain reaction amplification using 20 arbitrary decamers as primers. Out of these primers, 4 generated 17 highly reproducible RAPD markers, producing almost six polymorphic bands per primers. Four of 6 primers tested generated amplified fragments which were polymorphic between different individuals. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from individuals. Rainbow trout was distinctly observed 3 specific DNA markers (2. 3, 2.0 and 1.3kb) in bandsharing. Individual fragments generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in rainbow trout sperm DNA. The RAPD polymorphism generated by this primer may be used as a genetic marker for individual identification The RAPD-PCR technique has been shown to reveal informative polymorphism in many species of fish. The present results demonstrate that RAPD markers are abundant, reproducible and provide a basis for future gene mapping and MAS in these important aquaculture species using RAPD polymorphic markers. It is concluded that RAPD polymorphisms are useful as genetic markers for fish breed differentiation.

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Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

  • Byoung-Seon Yang
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.183-187
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    • 2003
  • Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated, Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type lb and 15 strains from Chungnam University Hospital to RAPD type I or II. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

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Molecular Typing of Leuconostoc citreum Strains Isolated from Korean Fermented Foods Using a Random Amplified Polymorphic DNA Marker

  • Kaur, Jasmine;Lee, Sulhee;Sharma, Anshul;Park, Young-Seo
    • Food Engineering Progress
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    • v.21 no.2
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    • pp.174-179
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    • 2017
  • For preliminary molecular typing, PCR-based fingerprinting using random amplified polymorphic DNA (RAPD) is the method of choice. In this study, 14 bacterial strains were isolated from different Korean food sources, identified using 16S rRNA gene sequencing, and characterized through RAPD-PCR. Two PCR primers (239 and KAY3) generated a total of 130 RAPD bands, 14 distinct PCR profiles, 10 polymorphic bands, one monomorphic band, and four unique bands. Dendrogram-based analysis with primer 239 showed that all 14 strains could be divided into seven clades out of which clade VII had the maximum of seven. In contrast, dendrogram analysis with the primer KAY3 divided the 14 L. citreum strains into four clades out of which clade IV consisted of a maximum of 10 strains out of 14. This research identified and characterized bacterial populations associated with different Korean foods. The proposed RAPD-PCR method, based on sequence amplification, could easily identify and discriminate the lactic acid bacteria species at the strain-specific level and could be used as a highly reliable genomic fingerprinting tool.

Differences by RAPD-PCR Analysis within and between Rockfish (Sebastes schlegeli) Populations from the Yellow Sea and the Southern Sea in Korea (황해 및 남해산 조피볼락 (Sebastes schlegeli) 개체군 사이의 RAPD-PCR 분석에 의한 차이)

  • Yoon, Jong-Man;Kim, Jong-feon
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.359-369
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    • 2001
  • Polymerase chain reaction (PCR) amplification of DNA as 30 different arbitrary primers and random amplified polymorphic DNAs (RAPD) analysis were performed on genomic DNA extracted from the blood of the marine rockfish (Sebastes schlegeli) from the Yellow Sea and the Southern Sea. The unique properties of the genomic DNA were used to investigate the features of the population dynamics and origins of the species. Out of 30 primers, seven generated 207 highly reproducible RAPD polymorphic products, producing approximately 2.7 polymorphic bands per primer. About 67.4% of total amplified products (307) were either polymorphic (207) to rockfish. The degree of similarity varied from 0.22 to 0.63 as calculated by bandsharing analysis. Also, the average level of bandsharing was 0.39$\pm$0.02 within the rockfish strains. The electrophoretic analysis of RAPD-PCR products showed the relatively high levels if variation between different individuals in rockfish from the Yellow Sea. However, the RAPD outlines obtained with DNA of different rockfish strains from the Yellow Sea and the Southern Sea in Korea were very similar. Also, a small number of polymorphic bands were identified. Even if further analyses or more rockfish populations are required, this result implies RAPD analysis reflects genetic differences between the geographical strains of the rockfish.

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Construction of a Genetic Linkage Map of Shiitake Mushroom Lentinula Edodes Strain L-54

  • Hoi-Shan, Kwan;Hai-Lou, Xu
    • BMB Reports
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    • v.35 no.5
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    • pp.465-471
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    • 2002
  • From fruiting bodies of L. edodes strain L-54, single-spore isolates (SSIs) were collected. Two parental types of L-54 were regenerated via monokaryotization. By means of random-amplified polymorphic DNA (RAPD), DNA samples from L-54, its two parental types, and 32 SSIs were amplified with arbitrary primers. Dedikaryotization was demonstrated, and 91 RAPD-based molecular markers were generated. RAPD markers that were segregated at a 1:1 ratio were used to construct a linkage map of L. edodes. This RAPD-linkage map greatly enhanced the mapping of other inheritable and stable markers [such as those that are linked to a phenotype (the mating type), a known gene (priA) and a sequenced DNA fragment (MAT)] with the aid of mating tests, bulked-segregant analysis, and PCR-single-strand conformational polymorphism. These markers comprised a genetic map of L. edodes with 14 linkage groups and a total length of 622.4 cM.

RAPD-PCR Analysis in Fusarium species (Fusarium 종에서의 RAPD-PCR분석)

  • 민병례;양연주;최영길
    • Korean Journal of Microbiology
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    • v.35 no.2
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    • pp.107-114
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    • 1999
  • To assess genetic diversity amoug 21 strains from sixleen Frrsn~i~nn species , we used RAPD(rando1n amplified pol.ymorphic DNA) analysis based on PCR(po1ymerase chain reaction). Eleven primers showing Ule polymorphism were chosen from the 40 random pnmers-tcstcd. A total of 263 polymorphic bands were generated by the primers and the size of amplified DNA fragments ranged from 0.1 lo 3.0 kb. Sirnilku-it), coefficients between strains were calcnlatcd, and UPGMA cluster analysis was used to generate a dendrogram showing relationships among them. The results from RAPD-PCR analysis were grouped into four main groups at the si~nilarity level of 0.627.

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Differentiation of Lentinus edodes Isolates in Korea by Isozyme Polymorphisms and Random Amplified Polymorphic DNA (RAPD) Analysis (Isozyme Polymorphism 및 Random Amplified Polymorphic DNA(RAPD) Pattern에 의한 표고 버섯 품종간 비교)

  • Park, Won-Mok;Ko, Han-Gyu;Park, Ro-Jo;Hong, Ki-Sung;Kim, Gyu-Hyun
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.176-190
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    • 1997
  • Sixty-three isolates of Lentinus edodes obtained from Korea were used to assess the genetic similarity by isozyme polymorphisms and random amplified polymorphic DNA patterns. The activities of esterase, peroxidase and acid phosphatase displayed 10, 7 and 3 distinct isozyme patterns, respectively. By combining the isozyme patterns obtained with the 3 enzymes, every isolate showed its own distinct electrophoretic phenotypes. A distance matrix calculated between all pairs of 63 electrophoretic phenotypes based on the presence or abscence of isozyme bands were analyzed by the group-average method. Results of the cluster analysis assinged the 63 phenotypes into six major groups. In the analysis of random amplified polymorphic DNA patterns, all isolates of Lentinus edodes were devided into five RAPD groups.

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