• Title/Summary/Keyword: RAPD technique

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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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RAPD Analysis of DNA Polymorphism and Genetic Species-Specificity Using PCR Technique in the Marine Microalgae

  • Yoon, Jong-Man;Chang, Kye-Nam
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.487-488
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    • 2000
  • Genomic DNA was isolated from the marine microalgae representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. The electrophoretc analysis of PCR-RAPD products showed hig levels of variation between different genus and little variation between different species. Outer of these primers, 6 generated 248 highly reproducible RAPD markers, producing almost seven polymorphic bands per primers. The degree of similarity frequency between Chaetoceros gracilis and Chaetoceros calcitrans species showed 90% as calculated by sharing analysis. The RAPD polymorphism generated by this primer may be used as a genetic marker for genus or species identification in important marine microalgae. (omitted)

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Assessment of Genetic Variability in Two North Indian Buffalo Breeds Using Random Amplified Polymorphic DNA (RAPD) Markers

  • Sodhi, M.;Mukesh, M.;Anand, A.;Bhatia, S.;Mishra, B.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1234-1239
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    • 2006
  • Murrah and NiliRavi are the important North Indian buffalo breeds occupying the prominent position of being the highest milk producers. These breeds are more or less similar at morphological as well as physiological levels. The technique of RAPD-PCR was applied in the present study to identify a battery of suitable random primers to detect genetic polymorphism, elucidation of the genetic structure and rapid assessment of the differences in the genetic composition of these two breeds. A total of 50 random primers were screened in 24 animals each of Murrah and NiliRavi buffaloes to generate RAPD patterns. Of these, 26 (52%) primers amplified the buffalo genome generating 263 reproducible bands. The number of polymorphic bands for the 26 chosen RAPD primers varied from 3 (OPG 06 and B4) to 26 (OPJ 04) with an average of 10.1 bands per primer and size range of 0.2 to 3.2 kb. DNA was also pooled and analyzed to search for population specific markers. Two breed specific RAPD alleles were observed in each of Murrah (OPA02 and OPG16) and NiliRavi (OPG09) DNA pools. RAPD profiles revealed that 11 (4.2%) bands were common to all the 48 individuals of Murrah and NiliRavi buffaloes. Pair-wise band sharing calculated among the individual animals indicated considerable homogeneity of individuals within the breeds. Within breed, band sharing values were relatively greater than those of interbreed values. The low genetic distance (Nei's) value (0.109) estimated in this study is in accordance with the origin and geographical distribution of these breeds. The RAPD analysis indicated high level of genetic similarity between these two important North Indian buffalo breeds.

Morphological and Molecular Characterization of Thamnocalamus falconeri Hook f. ex. Munro

  • Tiwari, Chandrakant;Bakshi, Meena;Nautiyal, Subhash
    • Journal of Forest and Environmental Science
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    • v.31 no.3
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    • pp.214-224
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    • 2015
  • The economy of India and so also of many Asian countries depends on bamboos and their uses are not only in domestic items but also in rural housing and raw materials to several industries and germplasm characterization is an important link between the conservation and utilization of plant genetic resources. Classical taxonomic studies of the bamboos are based on floral morphology and growth habit, which can cause problems in identification due to erratic flowering coupled with different biotic agencies and environmental factors. Identification and genetic relationships among accessions of Thamnocalamus falconeri were investigated using morphology and random amplified polymorphic DNAs (RAPD) technique. Analysis started by using 51 vegetative characters and forty two 10-mer primers that allowed us to distinguish different genotypes hailing from different eco- zones of Garhwal Himalayas (India). The selected primers (12) were used for identification and for establishing a profiling system to estimate genetic diversity. A total of 79.33% polymorphism was estimated by using 12 selected primers. The genetic similar analysis was conducted based on binary digits i.e. presence (1) or absence (0) of bands, which revealed a wide range of variability among the species whereas genetic relatedness was quite high based on vegetative characters. Cluster analysis clearly showed two major clusters for both of the markers viz. morphology and RAPD belonging to 10 accessions of T. falconeri. Two major clusters were further divided into minor clusters. Cluster based on RAPD marker showed grouping of accessions of closed locality whereas analogy was reported for vegetative traits. The RAPD technique has the potential for use in species identification and genetic relationships studies of bamboo for breeding program.

Analysis of Genetic Variability Using RAPD Markers in Paeonia spp. Grown in Korea

  • Lim, Mi Young;Jana, Sonali;Sivanesan, Iyyakkannu;Park, Hyun Rho;Hwang, Ji Hyun;Park, Young Hoon;Jeong, Byoung Ryong
    • Horticultural Science & Technology
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    • v.31 no.3
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    • pp.322-327
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    • 2013
  • The genetic diversity and phylogenetic relationships of eleven herbaceous peonies grown in Korea were analyzed by random amplified polymorphic DNA (RAPD). Twenty-four decamer RAPD primers were used in a comparative analysis of these Korean peony species. Of the 142 total RAPD fragments amplified, 124 (87.3%) were found to be polymorphic. The remaining 18 fragments were found to be monomorphic (12.7%) shared by individuals of all 11 peony species. Cluster analysis based on the presence or absence of bands was performed by Jaccard's similarity coefficient, based on Unweighted Pair Group Method with Arithmetic Averages. Genetic similarity range was 0.39 to 0.90 with a mean of 0.64. This study offered a rapid and reliable method for the estimation of variability among different peony species which could be utilized by the breeders for further improvement of the local peony species. Also, the results propose that the RAPD marker technique is a useful tool for evaluation of genetic diversity and relationship amongst different peony species.

Morphological Characteristics and Genetic Diversity Using the RAPD Technique in the Arksllell, Scapharca broughtosnii (Schrenck) Sronn Korea and China (한국산 및 중국산 피조개, Scapharca broughtonii(Schrenck)의 형태적 특성과 RAPD 기법을 이용한 유전적 분석)

  • LEE Jeong Mee;PARK Ji-Won;YOO Myung Suk;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.30 no.2
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    • pp.297-304
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    • 1997
  • Morphological and Benetic comparision were studied for the arkshell populations of Korea and China. The shell was collected from commercial arkshell bed in Jinhae Bay, Korea and from an hatchery in Young Seong, China. Shell Parameters, number of radial ribs and random amplified polymorphic DNA (RAPD) band were measured. The two populations had morphological and genetic differences. There was significant morphological difference $(P\leq0.05)$ in the ratio of the longest radial rib to the shell length. Posterior parts of the Chinese arkshell were more elongated than those of the Korean arkshell. Number of radial ribs were $36\~41$ for the Korean arkshell and $39\~43$ for the Chinese arkshell. RAPD markers generated tty each of 19 primers were $2\~6$ bands. Genetic diversity between the two Populations was clear since the genetic similarity was very low, not exceeding 0.29. The genetic similarity among the Korean arkshells $(0.58\~0.40)$ was higher than that of the Chinese arkshells $(0.48\~0.32)$.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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Classification of Lilium Using Random Amplified Polymorphic DNA (RAPD) (Random Amplified Polymorphic DNA(RAPD)를 이용한 나리(Lilium )의 품종구분)

  • Choi, Hae-Sun;Kim, Kyung-Su;Choi, Jang-Kyung;Lee, Kyung-Kook;Hong, Dae-Ki;Kang, Won-Hee;Lee, Youn-Su
    • Horticultural Science & Technology
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    • v.17 no.2
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    • pp.144-147
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    • 1999
  • RAPD technique was employed for the genetic analysis of major Lilium cultivars and horticultural hybrids. As a result of RAPD with 10-mer random primers, total 107 bands were observed within 300bp and 2kb range, and the same band patterns were observed within the same cultivar for different primers. However, Casa Blanca in Orientals and Adelina in Asiatics showed different band patterns with others in the same. Cultivars within L. longiflorum showed different band patterns. RAPD markers produced with random primers OPA- 02, 03, 04, 14, 16 and 17 can be used for the classification of Lilium cultivars.

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