• Title/Summary/Keyword: polymorphic bands

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Genetic Polymorphism of Marsh Clam (Corbicula leana) Identified by RAPD- PCR

  • Yoon Jong-Man;Park Kwan-Ha;Choe Sun-Nam
    • Fisheries and Aquatic Sciences
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    • v.6 no.1
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    • pp.13-19
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    • 2003
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang, Muan and a Chinese site was extracted to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction (RAPD- PCR). Out of 20 primers, seven primers produced amplified fragments which were consistently polymorphic. A total of 1,246 amplified products were produced of which 530 were polymorphic $(42.5\%)$. The number of polymorphic bands produced per primer varied from 40 to 122 with an average of 75.7 in marsh clam from Gochang. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic. Also, about $4.34\%$ of total polymorphic bands were specific to marsh clam from Gochang. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, which were polymorphic. This common bands in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of 12 specific bands. The intra-population variation was revealed in the band patterns identified by this primer. The random primer OPB-12 (CCTTGACGCA) yielded the amplified fragments which were consistently polymorphic between the marsh clams from Gochang and from Muan. This primer produced a total of 77 polymorphic bands: 31 bands from Gochang, 14 from Muan and 32 from the Chinese populations. An average of polymorphic bands were 1.8 from Gochang and 2.5 from the Chinese populations. This value obtained from the Chinese population was higher than those from the two domestic populations. Generally, the RAPD polymorphism generated by these primers may be useful as a genetic marker for strain or population identification of marsh clam.

Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Hwan-ha;Choe, Sun-Nam
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2002.05a
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    • pp.279-280
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana) from Gochang was extracted in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and/or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35 kb) identifying individuals was observed in lane 22.

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Genetic Similarity and Difference of Marsh Clam (Corbicula leana) Obtained by RAPD-PCR

  • Yoon, Jong-Man;Park, Kwan-Ha;Choe, Sun-Nam
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2002.08a
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    • pp.171-172
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    • 2002
  • Genomic DNA from the muscle of marsh clam (Corbicula leana)from Gochang was extrected in order to identify genetic differences and similarity by randomly amplified polymorphic DNAs-polymerase chain reaction. 3.28 of the 23.0 polymorphic bands per lane were found to be polymorphic in marsh clam. Also, about 4.34% of total polymorphic bands were either specific to marsh clam. The major common bands of 0.28 kb generated by primer OPB-15 (GGAGGGTGTT) were present in every individuals, respectively, which were polymorphic. This common bands which present in every individuals should be diagnostic of specific strains, species and-or their relatedness. Primer OPB-19 (ACCCCCGAAG) produced the highest number of specific bands, which was 12. The specific minor band of 0.07 kb was present in lane 22, which were polymorphic. Especially, only a specific band (1.35kg) identifying individuals was observed in lane 22.

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Identification of Different Species and Dultivars of Brassica by SDS-PAGE, Isozyme and Molecular Marker

  • Mukhlesur Rahman Md.;Hirata Yutaka
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.27-35
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    • 2005
  • Eighty-five different cultivars of Brassica rapa, B. juncea, B. nap us, B. carinata, B. oleracea and hexaploid Brassica collected from Bangladesh, Japan, China and Denmark were analyzed by SDS-PAGE for seed and leaf protein variations, using esterase, acid phosphatase and peroxidase isozyme analysis. Ten polymorphic bands were identified from seed protein however no identifiable polymorphic band was found in the leaf protein. Polymorphic markers clearly distinguished the different Brassica species as well as yellow sarson (YS) and brown seeded (BS) cultivars of B. rapa. The $F_1$ cross between YS and brown seeded cultivars showed the existance of all poly-morphic bands of the respective parents. The Bangla-deshi and Japanese cultivars of B. rapa differed in the amount of seed protein. In the case of isozyme analysis, esterase showed the highest number of polymorphic bands (13) followed by acid phosphatase (9) and peroxidase (5). These polymorphic markers were very effec-tive for classification of all the species studied in this experiment. In parentage tests using isozymes, the hybridity of intra-and-interspecific crosses of almost all the seedlings could be identified from their respective cross combinations. Esterase polymorphism showed a clear differentiation between YS and BS types of B. rapa. In addition, two esterase polymorphic markers were iden ified to differentiate some cultivars of B. juncea. Segregation patterns in these two esterase bands showed a simple Mendelian monohybrid ratio of 3:1 in $F_2$, 1:1 in test cross and 1:0 in back cross progenies. No polymorphic band was identified to distinguish different cultivars of the same species by acid phosphatase or peroxidase. Polymerase Chain Reaction (PCR) was carried out with seed coat color specific marker of B. juncea. The yellow seeded cultivars produced a strong band at 0.5 kb and weak band 1.2 kb. In the addition of these two specific bands, Japanese yellow-seeded cultivars expressed two more weak bands at 1.0 kb and 1.1 kb. Where the brown seeded cultivars generated a single strong band at 1.1 kb. In segregating population, the yellow seed coat color marker segregated at a ratio 15 (brown) : 1 (yellow), indicating the digenic inheritance pattern of the trait.

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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Genetic Similarity and Variation in the Cultured and Wild Crucian Carp (Carassius carassius) Estimated with Random Amplified Polymorphic DNA

  • Yoon, Jong-Man;Park, Hong-Yang
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.4
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    • pp.470-476
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    • 2002
  • Random amplified polymorphic DNA (RAPD) analysis based on numerous polymorphic bands have been used to investigate genetic similarity and diversity among and within two cultured and wild populations represented by the species crucian carp (Carassius carassius). From RAPD analysis using five primers, a total of 442 polymorphic bands were obtained in the two populations and 273 were found to be specific to a wild population. 169 polymorphic bands were also produced in wild and cultured population. According to RAPD-based estimates, the average number of polymorphic bands in the wild population was approximately 1.5 times as diverse as that in cultured. The average number of polymorphic bands in each population was found to be different and was higher in the wild than in the cultured population. Comparison of banding patterns in the cultured and wild populations revealed substantial differences supporting a previous assessment that the populations may have been subjected to a long period of geographical isolation from each other. The values in wild population altered from 0.21 to 0.51 as calculated by bandsharing analysis. Also, the average level of bandsharing values was $0.40{\pm}0.05 $ in the wild population, compared to $0.69{\pm}0.08$ in the cultured. With reference to bandsharing values and banding patterns, the wild population was considerably more diverse than the cultured. Knowledge of the genetic diversity of crucian carp could help in formulating more effective strategies for managing this aquacultural fish species and also in evaluating the potential genetic effects induced by hatchery operations.

Genetic Polymorphism among Korean Salmonids Determined by RAPD (Randomly Amplified Polymorphic DNA) Analysis

  • Park, Jung-Youn;Kim, Mi-Jung
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.2
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    • pp.102-111
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    • 2007
  • RAPD analyses using 60 OPERON primers and 13 URPs were performed in order to assess the genetic variation and frequency of polymorphisms in Korean salmonids. RAPDS were very reproducible and most useful at the sub-species level. In RAPD analysis, 138 polymorphic bands were detected between Oncorhynchus masou subspecies and 99 bands were generated in two types of rainbow trout. Estimated genetic distances between O. masou subspecies were 0.28794, and between wild rainbow trout and an albino mutant was 0.22786. Each species of salmonid was well characterized using URP 4R, the obtained bands could be useful as a species specific RAPD markers.

Genetic Diversity Evaluation of Thamnocalamus spathiflorus (Trin.) Munro Accessions through Morphological and Randomly Amplified Polymorphic DNA (RAPD) Markers

  • Tiwari, Chandrakant;Bakshi, Meena;Gupta, Dinesh
    • Journal of Forest and Environmental Science
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    • v.35 no.2
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    • pp.90-101
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    • 2019
  • Biodiversity refers to the total number and variation among species of flora and fauna of an area. Due to tremendous biotic especially anthropogenic pressure these natural resources are being vanishing. In present study genetic diversity among accessions of Thamnocalamus spathiflorus was evaluated. A total of 51 vegetative characters and 42 primers (10-mer) were screened. Out of 42 screened primers, 28 polymorphic primers were selected for further analysis. A total of 263 bands were recorded as polymorphic whereas 48 bands were monomorphic. The resolving power (Rp) of 28 Randomly Amplified Polymorphic DNA (RAPD) primers ranged from 4.6 (OPE08) to 17.6 (OPA11). The polymorphic information content (PIC) value ranged from 0.21 (OPAH09) to 0.44 (OPG02). The result revealed high degree of genetic relatedness (56 to 80%). Cluster analysis revealed two major clusters both for morphology as well as RAPD. Unlike morphological characterization, the accession (D5) from Bahli, Rampur, Shimla (H.P.) was clustered separately from the others in RAPD cluster analysis. Accessions with closed locality grouped together through RAPD marker system however analogy was recorded for morphological traits. The study conducted reflects the utility of RAPD technique for species identification and phylogenetic studies in bamboo for conducting bamboo breeding program.

Application of RAPD Methods in Meat for Beef Breed Identification

  • Choy, Y.H.;Oh, S.J.;Kang, J.O.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1655-1658
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    • 2001
  • Bovine genome samples were collected from meat of three different beef breeds (Hanwoo, Holstein and imported beef breed) that are commercially merchandized in Korean beef market. Operon B (OPB)-kits were used as random primers (3, 7, 10, 11, 12, 14) in random amplified polymorphic DNA (RAPD) method on whole genome. Each primer provided characteristic bands that were highly polymorphic. Each single primer could provide relatively efficient polymorphic band patterns among breeds. However, use of two or more primers in combination is recommended to improve resolution of experiments with higher molecular weight bands of DNA. In our experiments, OPB-11 resolved well between beef cattle breeds and Holstein. And OPB-7, 12 and 14 could be combined with OPB-11 to identify Hanwoo beef from the other two kinds of beef.

Genetic Polymorphisms and phylogenetic Relationships of Italian Ryegrass Cultivars Based on Random Amplified Polymorphic DNA ( RAPD ) Markers (RAPD 표지인자를 이용한 이탈리안 라이그라스 품종의 유전적 변이 및 유연관계 분석)

  • 임용우;이승재;신정섭;정영수;최기준;임영철;임근발;박병훈
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.1
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    • pp.35-42
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    • 1998
  • Eleven Italian ryegrass cultivars were examined for their genetic polymorphisms and phylogenetic relationships using randomly amplified polymorphic DNA (RAPD) markers. In RAPD analysis of 34 random primers, 96 of total 162 bands obtained from 16 primers were polymorphic and sizes of polymorphic band ranged between 0.5 and 1.5kb. Number of bands amplified per primer was varied from 3 to 16 and average number was 14.8. Phylogenetic relationship among cultivars based on the RAPD analysis was examined using UPGMA computer program. In pairwise genetic similarity test of 11 Italian ryegrass cultivars, Grazer and Orlando showed highest coefficient of genetic similarity as 0.740, whereas Marshall and Orlando was lowest as 0.438. Eleven Italian ryegrass cultivars were grouped into 3 major clusters and genetic distance of clusters ranged between 0.567 and 0.646, indicating low level of genetic variation.

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