• Title/Summary/Keyword: RAPD method

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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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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.

Intraspecific Diversity of Korean Takydromus wolteri(Reptilia: Squamata) Based on Randomly Amplified Polymorphic DNA (RAPD) Analysis (RAPD를 이용한 한국산 줄장지뱀(Reptilia: Squamata)의 종내 다양성에 관한 연구)

  • 장민호;송재영;정규회
    • Korean Journal of Environmental Biology
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    • v.22 no.2
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    • pp.295-299
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    • 2004
  • The lacertid lizard Takydromus wolteri widely distributed in South Korea, but intraspecific diversity of this species was not almost studied. T. wolteri represented by 8 specimens collected from 5 localities were analyzed by the Randomly Amplified Polymorphic DNA (RAPD) method with 28 random decamer primers and the phenogram constructed by using the Unweighted Pair Group Method with Arithmetic Mean (UPGMA) method based on RAPD data. The populations of T. wolteri using in this paper were collected from Gyeonggi-do, Chungcheongbuk-do, Jeju-do, Jeollanam-do and Gyeongsangnam -do in South Korea. Among 68 different bands detected in RAPD analysis, 59 bands (87%) showed polymorphism. The phenogram showed that the populations of T. wolteri were grouped into two. These results suggest that T. wolteri were supported intraspecific diversity.

Identification study of rare and high-priced natural products used for oriental medicine by RAPD analysis (RAPD 분석법을 이용한 산삼, 웅담, 녹용 등의 한약재 판별연구)

  • Cho, Dong-Wuk
    • Korean Journal of Oriental Medicine
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    • v.1 no.1
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    • pp.471-476
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    • 1995
  • Natural products used for oriental medicine often come from various geographical sources, after several different distribution channels. Therefore some form of quality control procedure is required to safeguard naturl products for prescriptions purposes. To achieve this, systematic apprroaches such as morphological examination, microscopic analysis of powdered herbs and chemical analysis can be carried out. However, to ensure absolute criteria for quality assurance of natural products, DNA fingerprinting method such as RAPD(Random amplified polymorphism DNA) analysis can be used for authentication of natural products for authenticatin of natural products. In this study, warious oligonucleotide primers will be synthesized for the detection of RAPD markers and also parameters of affecting PCR(Polymerase Chain Reaction) in the detection of RAPD markers of rare and high-priced natural products will be studied with genomic DNA of chosen samples.

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Discrimination and Genetic Relationship of Adenophorae triphylla(Thunb) A.DC. var. japonica Hara and Codonopsis lanceolata Trauty using RAPD analysis (RAPD분석에 의한 잔대와 더덕의 유연관계 비교 및 감별)

  • Lee, Mi-Young;Mo, Suk-Yeon;Kim, Du-Whan;Oh, Seong-Eun;Ko, Byoung-Seob
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.3
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    • pp.205-210
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    • 2001
  • Dried parts of the two species are difficult to distinguish morphologically, thus Codonopsis radix has been sold instead of Adenophorae radix in herbal medicine market. Therefore, this study was conducted to develop the genetic marker through the examination of the phylogenetic relationships between two Adenophora triphylla(Thunb.) A. DC. var. japonica Hara, two Adenophora radiatifolia Nakai, five Codonopsis lanceolata(Sieb. et Zucc)Trautv. using RAPD analysis. Fifty decarmer oligonucleotide primers were screened for the RAPD analysis, and four primers generated distinct RAPD markers specific to Adenophorae radix and Codonopsis radix. Based on the RAPD patterns, the genetic relationships between three herbal medicine were analyzed by UPGMA method. As a result, Adenophorae radix and Codonopsis radix were classified into two major subgroups on the basis of the genetic similarity coefficient. The specific RAPD patterns generated by the selected primers were reproducible from dried materials. Furthermore, the specific RAPD patterns were produced from the mixture of dried roots of A. triphylla and C. lanceolata. These results prone the usefulness of the RAPD analysis for the discrimination of pure materials from the mixtures of A. triphylla and C. lanceolata.

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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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RAPD Analysis on the Species of Pinelliae Tuber (RAPD 방법을 이용한 반하류 한약재의 감별 연구)

  • 배명효;김규열;정유헌;최호영
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.16-22
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    • 2000
  • This study intends to report the significance of several experimental results obtained from analysing the genes extracted from the plants and herbal medicine such as P. temalta (Thunb.) Breit, A. amurense var serratum Nakai, A. erubescens (Wall.) Schott, Pinelliae Tuber and Arisaematis Tuber, mainly by the method of RAPD(randomly amplified polymorphic DNA) and the method of RFLP(restriction fragment length polymorphism) on ITS(internal transcribed spacer) region. Genomic DNA could be extracted from both original plants and dried materials. DNA fragments of P. temata kind and A. amurense kind showed the same aspect separately within the same species under the method of RAPD using random primer, while various aspects(polymorphism) were discovered among different species. In RAPD analysis by uniprimer, common bands were extracted from all types of P. temata in the case of uniprimer #4, which were distinguished from the kind of A. amurense. Other polymorphic bands appeared in between different A. amurense species as well. In the case of uniprimer #11, particular band came out in the kind of P. temata. On the other hand, in the case of uniprimer #5, #6, and #8, various bands(polymorhism) were revealed in both kinds of P. temata and A. amurense. Although further study is needed to ascertain whether these results are due to the differences of species, kinds, or growing place, the results could be used as a scientific method of identifying the substitutes for A. amurense genus. The author believes that as if P. temata class of plants used in this experiment are different among themselves in terms of the shape, size and property, those are clearly a class of P. temata or belong to the same genus.

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Genetic Diversity and Discrimination of Astragalus Membranaceus Bunge and A. Membranaceus var. Mogholicus Using RAPD Markers (RAPD마커를 이용한 황기의 유전적 다양성 및 기원판별)

  • Bang Kyang Hwan;Huh Man Kyu;Cho Jean Hyeang
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.825-829
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    • 2004
  • This study was carried out to differentiate the origins of Astragalus membranaceus Bunge and A. membranaceus Bunge var. mogholicus Nakai. To identify the variation of the RAPD patterns between domestic and foreign Astragalus species, 40 random primers were applied to ten accessions of A. membranaceus and six accessions of A. membranaceus var. mogholicus genomic DNA, respectively, Ten primers of 40 primers could be used to discriminate the origins and 33 polymorph isms among 44 scored DNA fragments (33 fragments are specific for A. membranaceus and A. membranaceus var. mogholicus) were generated using these primers, 75.0 % of which were polymorphic. Especially, three primers of ten primers, OPA17, OPA11 and OPB11, were useful to differentiate between domestic and foreign Astragalus species. RAPD data from the 10 primers were used for cluster analysis and cluster analysis of RAPD markers showed that the two groups are distinct genetically. Consequently, RAPD analysis was a useful method to discriminate between A. membranaceus and A. membranaceus var. mogholicus.

Random Amplified Polymorphic DNA Analysis for Typing Extended-Spectrum-β-Lactamase of Klebsiella pneumoniae

  • Yang, Byoung-Seon
    • Korean Journal of Clinical Laboratory Science
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    • v.37 no.3
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    • pp.149-154
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    • 2005
  • Fifty-one extended-spectrum-${\beta}$-lactamase(ESBL) producing Klebsiella pneumoniae strains were isolated from national university hospitals. All K. pneumoniae strains showed resistance to broad-spectrum antibiotic and most of them presented resistance to amikacin, gentamicin and ciprofloxacin. The results of amplified polymorphic DNA (RAPD) pattern for randomly isolated fifty-one strains were as follows; both twenty-one strains from Chungnam National University hospital and ten strains from Chungbuk National University hospital showed RAPD type Ia and Ib. However, twenty strains isolated from Gyeongsang National University hospital belonged to RAPD type IIa and IIb. All isolates were divided into four molecular types and showed high level of genetic diversity. These results suggested that RAPD analysis provided a rapid and simple method for analysing genotypes of ESBL.

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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.