• Title/Summary/Keyword: RAPD

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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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A Study on Genetic Relationship between Sasang Constitutions by the Polymorphic Analysis of RAPD (RAPD 다형성 분석을 통한 사상체질간 유전적 상관관계에 관한 연구)

  • 이휘철;조동욱;조중호;서영우;이창수
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.62-68
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    • 2000
  • This study was carried out to establish genetic understanding of three Sasang constitutions of Taeumin, Soeumin and Soyangin by Random Amplified Polymorphic DNA(RAPD) analysis. We have applied RAPD analysis to pooled DNA sample as a means to achieve rapid screening of large numbers of primers for their capacity to reveal constitutions-specific polymorphisms. From an initial 440 primers, 13 polymorphic primers between different constitutions were selected. Bandsharing(BS) and mean average percentage difference(MAPD) calculated within and between three constitutions using RAPD fingerprint data showed a higher degree of homogenity within than between the constitutions and indicated measurable divergence between three constitutions. The RAPD bandsharing(BS) values ranged from 0.71 to 0.73 between the three constitutions. The interconstitution divergence was narrower between Taeumin and Soeumin, than between the other paired constitution comparisons. The genetic distance between the three constitutions was measured by BS values. Genetic distance by RAPD analysis was 0.007 between Taeumin and Soeumin, and 0.014 between Soyang and the others. In conclusion, the genetic distance of Teaumin and Soumin was closer than that of Soyangin in the analysis of RAPD by using 440 primers.

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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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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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Optimization of RAPD-PCR Conditions for Onions, Allium cepa L. (양파(Allium cepa L)의 RAPD 분석조건 최적화에 관한 연구)

  • 정순재;양보경;김익수;박선희;서전규;남재성;김현경;김도훈
    • Journal of Life Science
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    • v.10 no.2
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    • pp.182-187
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    • 2000
  • The optimized RAPD-PCR conditions, which can be utilized as a basic information for the analysis of the genetic characteristics were investigated with four onion varieties, named Changryungdaego, Yeoeuijuhwang, Yakwangju, and Dabonghwang using Operon primers, OPR01 (TGCGGGTCCT) and OPZ20 (ACTTTGGCGG). We tested several concentrations of DNA, primer, and MgCl2, annealing temperature, number of PCR cycle, and presence/absence of pre-heating time at the begining of PCR reation in the 25${mu}ell$ volume. The best RAPD profiles were obtained using 50ng of DNA, 5mM of primer, 1.5mM of MgCl2, 45$^{\circ}C$ of annealing temperature and an absence of pre-heating time. An establishment of the stable and reproducible RAPD-PCR conditions are expected to be useful for the subsequent RAPD-related investigation, such as genetic characterization of the onion strains, re-establishment of phylogenetic relationships and development of new varieties.

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Genetic Diversity of Phomopsis citri with Random Amplified Polymorphic DNA (RAPD) and Fungicide Resistance (RAPD 및 약제저항성을 이용한 감귤 검은점무늬병균의 유전적 다양성 분석)

  • 고영진;서정규;이태선;송장훈;권혁모;문덕영;문두길;한해룡
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.171-176
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    • 1998
  • Gentic diversity of 42 isolates of Phomopsis citri was analyzed with random amplified polymorphic DNA(RAPD) and fungicide resistance. RAPD profiles of genomic DNA of the isolates of P. citri and the degrees of their resistance to the fungicides mancozeb and propineb suggested the occurrence of genetic differentiation of P. citri distributed in Cheju. The isolates showed genetically diverse RAPD profiles according to the host species collected even from the same collection site and also according to the geographic origin collected even from the same host species. High levels of resistance to fungicides mancozeb and propineb were observed among the isolates of P. citri. However, there was no correlation between RAPD profiles of genomic DNA and levels of fungicide resistance of the isolates, suggesting that fungicide resistance of P. citri occurred irrespective of the host and geographic origin.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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Differentiation of Fusarium oxysporum f. sp. fragariae Isolates by Random amplified Polymorphic DNA (RAPD) Analysis. (Random Amplified Polymorphic DNA(RAPD)를 이용한 딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 분류)

  • 현재욱;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.41-46
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    • 1996
  • 본 실험은 이병 딸기의 조직에서 분리 동정된 시들음병균(Fusarium oxysporum f. sp. fragariae) 균주들의 유?거 변이를 random amplified polymorphic DNA(RAPD) marker들을 이용하여 조사하였다. 총 24개의 딸기 시들음병 균주들의 DNA를 주형으로 하여 16개의 random 10-mer primer들을 사용하여 증폭시킨 결과 총 231개의 marker들을 이용하여 유전적 변이를 조사해 본 결과 크게 RAPD I과 RAPD II의 2개 그룹으로 나눌 수 있었다. RAPD I그룹에 속하는 균주는 VCG A에 속하는 Y1, K1, K2, K3, K4, N2, N3, N4-1, N6-1, N6-2, N8, N9, N10, M1-2-1 균주, VCG B에 속하는 M4-1 균주 그리고 VCG C에 속하는 N1, Y2 균주들이었고, RAPD II그룹에는 VCG B에 속하는 M1-1, M2-2-1, M2-4-2, M3-2, M3-3-2 균주와 VCG D에 속하는 N1 1 균주가 속하였다. 이들 2그룹 간에는 31%의 유사성이 있었다.

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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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