• Title/Summary/Keyword: random amplification of polymorphic DNA(RAPD)

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Primers for typing Listeria spp. using Random Amplified Polymorphic DNA (RAPD) ANalysis (Listeria spp.의 RAPD typing을 위한 Primer의 분리력 비교)

  • 임형근;홍종해;박경진;최원상
    • Journal of Food Hygiene and Safety
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    • v.18 no.2
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    • pp.67-72
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    • 2003
  • Random amplified Polymorphic DNA (RAPD) analysis Is based on the amplification of random DNA segment using a single arbitrary primer. Polymorphic DNA patterns identified by this method can be used for typing Listeria monocytogenes. To select the primers for RAPD typing Listeria spp., the performance of 31 primers were compared by analyzing 13 Listeria spp. reference strains. Reproducible electrophoresis patterns were obtained. Among 31 primers, 6 primers (primer 6, HLWL74, UBC155, UBC127, Lis5, Lis11) showed better differentiation, when discrimination index, band clarity, band number, difficulty of band scoring were considered than the others. These primers will be useful far typing Listeria spp. in the future. Currently, we are under investigation for the RAPD typing of contaminated L. monocytogenes for the risk analysis of pork processing plant using these primers.

Classification of Listeria monocytogenes Isolates from Korean Domestic Foods Using Random Amplification of Polymorphic DNA and Serotyping Analysis (Random Amplification of Polymorphic DNA와 혈청학적 분석을 이용한 국내식품에서 분리한 Listeria monocytogenes의 분류)

  • Kim Hyun-Joong;Park Si-Hong;Kim Hae-Yeong
    • Microbiology and Biotechnology Letters
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    • v.34 no.1
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    • pp.23-27
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    • 2006
  • Molecular subtyping of Listeria monocytogenes, including type strains and isolates from Korean foods, were performed using random amplification of polymorphic DNA (RAPD). Each Listeria species showed specific RAPD band patterns, and L. monocytogenes serotypes and isolates were divided into two clusters. RAPD results showed that L. monocytogenes isolates from Korean foods were divided into two groups. Group I contained L. monocytogenes serotypes 1/2b and 4b, whereas Group II contained serotypes 1/2a and 1/2c. These results suggested RAPD as possible subtyping methods for Listeria species. Also, RAPD Results showed significant correlation between molecular subtyping and serotyping of L. monocytogenes, and classified two different groups of L. monocytogenes isolated from Korean foods.

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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Different RAPD patterns between Metagonimus yokogawai and Metagonimus Miyata type (RAPD분석을 이용한 요코가와 흡충과 미야타흡충의 분자생물학적 비교)

  • Yu, Jae-Ran;Jeong, Jin-Seong;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.35 no.4
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    • pp.295-298
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    • 1997
  • Genonlic DNA from Metagonimn vokogawci and Metagonimw Miyata type was amplified by polymerase chain reaction based on the random amplification of polymorphic DNA (RAPDI technique. Eight random 10-mer oligonucleotide primers (OPA-02, 5-TGCCGAGCTG-3; OPA-09, 5-GGGTAACGCC-3; OPA-17, 5-GTGATCGCAG-3; OPA-11, 5-CAATCGCCGT-3; OPA-13, 5-CAGCACCCAC-3; OPA-17. 5-GACCGCTrGT-3; OPA-19, 5-CAAACGTCGG-3; OPA-20, 5-GTTGCGATCC-3) WITH A G+C CONTENT FO 60-70% (Kit A. Operon Technologies Inc., California, USAI could produce distinguishable banding patterns between the two Metngonimus species. From the results of this study, it was suggested that Metcsonimus Miyata type has a different DNA sequence from M. WOkQgGUIGi. Key words: Metcgonimw vokognwai, MetnBonimw Miyata type, random amplification of polymorphic DNA (RAPD)

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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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Primers for Typing Salmonella spp. using Random Amplified Polymorphic DNA (RAPD) Analysis (Salmonella spp.의 RAPD Typing을 위한 Primer의 분리력 비교)

  • Lim, Hyung-Kum;Lee, Kyung-Hee;Hong, Chong-Hae;Park, Gyung-Jin;Choi, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.224-228
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis is based on the amplification of random DNA segment using a single arbitratrary primer. For typing Salmonella spp., polymorphic DNA patterns identified by this method can be used. To select the primers for RAPD typing Salmonella spp., the performances of 20 primers were compared by analyzing 16 Salmonella spp. reference strains. Reproducible electrophoresis patterns were obtained. Among the 20 primers tested, 4 primers (A, OPG04, OPG10, OPL03) showed better differentiation than the others. At the time discrimination index, band clarity, band number and difficulty of band scoring were considered. These primers will be useful for typing Salmonella spp. in the future. Curretly, we are under investigation for the RAPD typing of contaminated Slmonella spp. for the risk analysis of pork processing plant using the primers.

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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The Molecular Biological Marker in Bombyx mori and Spodoptera frugiperda Cells (Bombyx mori세포주와 Spodoptera frugiperda세포주의 분자생물학적 표식자)

  • Jin, Byeong-Rae;Je, Yeon-Ho;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • v.38 no.1
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    • pp.53-56
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    • 1996
  • To investigate the molecular biological marker in insect cells, BmN-4 and Sf-0 cells were analysed by SDS-PAGE and random amplification of polymorphic DNA. The results showed that the patterns of total cell protein and random amplification of polymorphic DNA were distinguished between BmN-4 and Sf-9 cells, suggesting that the unique major bands were useful as molecular biological marker in BmN-4 and Sf-9 cells.

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Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

Random Amplified Polymorphic DNA (RAPD) Identification of Genetic Variation in Chlorella species

  • CHO Jung Jong;KIM Yong-Tae;HUR Sung Bum;KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.761-769
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    • 1996
  • The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

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