• Title/Summary/Keyword: RAPD assay

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Fast Genetic Variation among Coliphage Quasispecies Revealed by a Random Amplified Polymorphic DNA (RAPD) Analysis

  • Kwon, Oh-Sik;Lee, Jae-Yung
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.166-171
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    • 1996
  • Genetic analysis was conducted on newly isolated coliphages form soil by using a RAPD assay. From the initial result, the coliphages were turned out to be different form one another but were closely related to .psi..lambda. due to the fact that they shared the samed RAPD maker in which other T phage testings failed to show. By using the primers EC01 or EC02, a fast genetic mutation of .psi.C1 was found by producing specific RAPD markers on the phages from the first filial progeny to the second filial progeny. When we made a RAPD assay with combined primers (EC01, EC05 and EC08), the genetic mutation was again confirmed in .psi.C1. The assay detection showed mutations in other coliphages such as .psi.C2 and .psi.C3 by revealing specific RAPD bands among different progeny phages, where genetic instability of the coliphages in implied.

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Classification of Korean Lentinula edodos Strains by Random Amplified Polymorphic DNA (RAPD) Markers (RAPD(Random Amplified Polymorphic DNA) 검정을 이용한 한국 표고균주의 계통분류)

  • Lee, Tai-Soo;Bak, Won-Chull;Kang, Ho-Duck;Kim, Se-Kwon;Byun, Byung-Ho;Yi, Chang-Keun;Lee, Won-Kyu;Min, Du-Sik
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.219-225
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    • 1997
  • Random Amplified Polymorphic DNA (RAPD) assay was used to identify seven typical Lentinula edodes (Berk.) Pegler strains isolated in Korea. Twenty primers from OPA-01 to OPA-20 were applied to generate the recognition of L. edodes strains. Out of 20 primers, nine primers showed efficient RAPD patterns to classify the 7 strains tested, but the rest eleven primers were not useful to be used. Even though there was no single primer that could classify all of the strains, any combination of two primers among the nine primers could identify the strains tested. Thus, RAPD assay turned out to be very precise method for classifying L. edodes strains.

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Phylogenetic Diversity and Comparison of RAPD Fingerprint Profile of Streptococcus iniae (Streptococcus iniae의 유전학적 다양성과 RAPD fingerprint profile의 비교)

  • Jeong Yong-Uk;Heo Moon-Soo
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.345-351
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    • 2006
  • Streptococcus sp. were isolated from cultured flounder (Paralichthys olivaceus) having Streptococcosis during 2004 to 2005 in Jeju Island. Ninety four Streptococcus iniae strains were isolated using biochemical test and multiplex PCR assay. Three genotypes (A, B, C-type) of S. iniae were appeared in the RAPD analysis and they showed international or local genetic polymorphism. Presently, S. iniae having A-type is a dominant S. iniae genotype in Jeju and showed band patterns at about 550, 850, 1000, 1300 and 2000 base pares. In this study, the reported P14 random primer, that used to distinguish serotypes of S. iniae could not be applied to distinguish Jeju island S. iniae's genetic polymorphism.

Aroclor 1254 May Induce Common DNA Effects in Developing Paralichthys olivaceus Embryos and Larvae

  • Min, Eun Young;Kang, Ju Chan
    • Fisheries and Aquatic Sciences
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    • v.17 no.4
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    • pp.461-469
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    • 2014
  • Polychlorinated biphenyls (PCBs) are persistent pollutants in aquatic environments, often causing the decline or disappearance of wild populations. In this study, we used a random amplified polymorphic DNA (RAPD) assay to evaluate the effects on the genomic DNA of olive flounder embryo and larval stages of exposure to Aroclor 1254 at concentrations of 1, 5, 10, 20, and $40{\mu}g/L$. We compared RAPD fingerprints of exposed and non-exposed samples. Polymorphisms were revealed as the presence and/or absence of DNA fragments between the two samples. A dose-dependent increase in the number of polymorphic bands was observed with Aroclor 1254 treatment. Also, RAPD profiles of animals exposed to Aroclor 1254 exhibited an increase in the frequency values (FV) compared to the control. A phenogram constructed using neighbor-joining method indicated that genomic template stability in developing embryo and larval stages was significantly affected at ${\geq}5{\mu}g/L$. This study suggested that DNA polymorphisms detected by RAPD analysis could be used as an investigative tool for environmental toxicology and as a useful biomarker in early life stages for the detection of potential genotoxicants.

Genetic polymorphism analysis of somatic embryo-derived plantlets of Cymbopogon flexuosus through RAPD assay

  • Bhattacharya, S.;Dey, T.;Bandopadhyay, T.K.;Ghosh, P.D.
    • Plant Biotechnology Reports
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    • v.2 no.4
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    • pp.245-252
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    • 2008
  • The genetic status of somatic embryo-derived plantlets of Cymbopogon flexuosus was examined by randomly amplified polymorphic DNA (RAPD) analysis. Auxins such as 2, 4-dichlorophenoxyacetic acid (2, 4-D) (1-4 mg/l) were used in Murashige and Skoog (MS) medium for induction of calli from rhizomatous explants of Cymbopogon flexuosus. Optimum calli were induced on MS medium supplemented with 2, 4-dichlorophenoxyacetic acid (2, 4-D) (3.5 mg/l) alone or in combination with $N^6-benzyladenine$ (2 mg/l). Somatic embryogenesis was achieved from long term calli when cultured on MS medium containing 2, 4-dichlorophenoxyacetic acid (2, 4-D) (2 mg/l) along with $N^6-benzyladenine$ (BA) (1-2 mg/l). Regeneration was achieved when freshly induced embryogenic calli were sub-cultured on MS medium supplemented with $N^6-benzyladenine$ (3 mg/l) alone. Long-term cultured embryos showed profuse minute rooting on regeneration medium supplemented with N6 -benzyladenine (3 mg/l). Microshoots were rooted in the presence of indole-butyric acid (IBA) (2 mg/l). DNA samples from the mother plant and 18 randomly selected regenerated plants from a single callus were subjected to RAPD analysis with 6 arbitrary decamer primers for the selection of putative somaclones. A total of 64 band positions were scored, out of which 19 RAPD bands were polymorphic. From genetic similarity coefficient based on RAPD band data sharing, it was found that the majority of the clones were almost identical or more than 92% similar to the mother plant, except CL2 and CL9 (66%) which showed highest degree of genetic change with CL2 and CL9 showing presence of two non-parental bands each.

Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers (해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계)

  • 김용국;진형주;박선미;진덕희;홍용기
    • 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.

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Identification and classification study of natural products by RAPD analysis (RAPD(Random Amplified Polymorphic DNA)법을 이용한 한약재의 판별 연구)

  • Kim, Dae-Weon;Kim, Do-Kyun;An, Sun-Kyong;Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.3 no.1
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    • pp.153-167
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    • 1997
  • Conventionally, identification and classification methods of natural products include the morphological survey and assay of chemical disposition, sing these methods, however, is not satisfying for the precise identification of natural products because they are often valiable in the compositions and morphology To standardize the natural products identification and classification, genomic DNA analysis such as RAPD, RFLP and Amp-FLP can be adopted for this purpose. In this study, various ginsengs and bear gall bladder were tested for the development of genetic identification and classification method. Varieties of ginsengs such as, P. ginseng, P. quinquefolium, P. japonicus and P. notoginseng, were genetically analyzed by RAPD. Also, DNA isolated from Bear blood and gall bladder, Ursus thibetanus, Ursus americanus and Ursus arctos, were analyzed by the same method. The results demonstrated that the identification and classification of bear gall bladder and various ginsengs were possible by RAPD analysis. Therefore, this method was thought to be used as a additional method for the identification and classification of other natural products.

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Relatedness Among Indiginous Members of Sclerotinia sclerotiorum by Mycelial Compatibility and RAPD Analysis in the Jordan Valley

  • Osofee, H.;Hameed, K.M.;Mahasneh, A.
    • The Plant Pathology Journal
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    • v.21 no.2
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    • pp.106-110
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    • 2005
  • Sclerotinia sclerotiorum attacks most of the vegetable crops in the Jordan valley. Twenty-five samples/isolates were obtained in a complete coverage of that region. They were characterized for their mycelium incompatibility, and specific gene amplified using the primer SSREV/SSFWD. All isolates gave similar single band around 278 bp. Thirteen isolates were completely incompatible with the other 12 ones. The latter ones fell into four subgroups of mycelium incompatibility. RAPD analysis using three primers (OPA-2, OPA-10, and OPA-18) clustered the 25 isolates into subgroups in agreement with their morphological separation, indicating close correlation between amplified gene(s) and the gene(s) of incompatibility. All highly virulent isolates were among the group of 13, indicating a well established genomic type pathogen in this region.