• Title/Summary/Keyword: Decamer primer

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Genetic Distances and Variations of Three Geographic Hairtail Populations Identified by PCR Analysis

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.18 no.3
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    • pp.167-172
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    • 2014
  • In the present study, muscle tissues were obtained separately from individuals from Atlantic hairtail population (AHP), Gunsan hairtail population (GHP) and Chinese hairtail population (CHP), respectively. The seven decamer primers were used to generate the shared loci, specific, unique shared loci to each population and shared loci by the three hairtail populations. Here, averagely, a decamer primer generated 64.7 amplified products per primer in the AHP population, 55.7 in GHP population and 56.4 in CHP population. The number of unique shared loci to each population and number of shared loci by the three populations generated by genetic analysis using 7 decamer primers in AHP, GHP and CHP population. 119 unique shared loci to each population, with an average of 17 per primer, were observed in the AHP population, and 28 loci, with an average of 4 per primer, were observed in the CHP population. The hierarchical dendrogram point out three main branches: cluster 1 (ATLANTIC 01 ~ ATLANTIC 07), cluster 2 (GUNSAN 08 ~ GUNSAN 14) and cluster 3 (CHINESE 15 ~ CHINESE 21). The shortest genetic distance displaying significant molecular difference was between individuals' CHINESE no. 16 and CHINESE no. 18 (0.045). In the long run, individual no. 01 of the AHP population was most distantly related to CHINESE no. 19 (genetic distance = 0.430). Consequently, PCR analysis generated on the genetic data displayed that the geographic AHP population was widely separated from CHP population, while individuals of CHP population were fairly closely related to those of GHP population.

Evaluation of Genetic Relationship and Fingerprinting of Rice Varieties using Microsatellite and RAPD Markers

  • Soo- Jin, Kwon;Sang-Nag, Ahn;Hae-Chune, Choi;Huhn-Pal, Moon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.2
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    • pp.112-116
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    • 1999
  • Genetic diversity of 31 rice varieties including 25 japonica and 6 indica varieties was evaluated using a combination of 19 microsatellite or simple sequence repeats (SSRs) and 28 random decamer oligonucle-otide primers. All 19 microsatellite primer sets representing 19 loci in the rice genome showed polymorphisms among the 31 varieties and revealed 91 alleles with an average of 4.80 bands per primer. Also all 28 random decamer primers used were informative and generated 114 non-redundant bands with a mean of 4.07 bands. Microsatellite markers detected higher number of alleles than random primers .although the mean difference was not statistically significant. A cluster analysis based on Nei's genetic distances calculated from the 205 bands resolved the 31 varieties into two major groups that correspond to indica and japonica subspecies, which is consistent with the genealogical information. As few as six random decamer primers or a combination of one microsatellite and four random decamer primers were sufficient to uniquely differentiate all 31 varieties. These combinations would be potentially useful in rice variety protection and identification considering that 25 out of 31 varieties used in this study are japonica rices with high grain quality and have close make up.

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Genetic Distances of Three Mollusk Species Investigated by PCR Analysis

  • Oh, Hyun;Yoon, Jong-Man
    • Development and Reproduction
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    • v.18 no.1
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    • pp.43-49
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    • 2014
  • Three species of Nortamea concinua (NC) and Haliotis discus hannai (HDH) from Tongyeong and Sulculus diversicolor supertexta (SDS) are widely distributed on the coast of the Yellow Sea, southern sea and Jeju Island in the Korean Peninsula under the innate ecosystem. There is a need to understand the genetic traits and composition of three mollusk species in order to evaluate exactly the patent genetic effect. PCR analysis was performed on DNA samples extracted from a total of 21 individuals using seven decamer oligonucleotides primers. Seven primers were shown to generate the unique shared loci to each species and shared loci by the three species which could be clearly scored. A hierarchical clustering tree was constructed using similarity matrices to generate a dendrogram, which was facilitated by the Systat version 10. 236 specific loci, with an average of 56.3 per primer, were identified in the NC species. 142 specific loci, with an average of 44.7 per primer, were identified in the HDH species. Especially, 126 numbers of shared loci by the three species, with an average of 18 per primer, were observed among the three species. Especially, the decamer primer BION-75 generated 7 unique loci to each species, which were identifying each species, in 700 bp NC species. Interestingly, the primer BION-50detected 42 shared loci by the three species, major and/or minor fragments of sizes 100 bp and 150 bp, respectively, which were identical in all samples. As regards average bandsharing value (BS) results, individuals from HDH species (0.772) exhibited higher bandsharing values than did individuals from NC species (0.655). In this study, the dendrogram obtained by the seven decamer primers indicates three genetic clusters: cluster 1 (CONCINNA 01~CONCINNA 07), cluster 2 (HANNAI 08~HANNAI 14), cluster 3 (SUPERTEXTA 15~SUPERTEXTA 21). Comparatively, individuals of HDH species were fairly closely related to that of SDS species, as shown in the hierarchical dendrogram of genetic distances.

Identification of RAPD markers linked to sex determination in guggal [Commiphora wightii (Arnott.)] Bhandari

  • Samantaray, Sanghamitra;Geetha, K.A.;Hidayath, K.P.;Maiti, Satyabrata
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.95-99
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    • 2010
  • Decamer RAPD primers were tested on dioeceious and hermaphrodite plants of Commiphora wightii to identify sex-specific molecular markers. Sixty different random decamer primers were screened out of which only three primers were found to be associated with sex expression. A ~1,280-bp fragment from the primer OPN06 was found to be present in all the female individuals. Another primer OPN 16 produced a unique ~400-bp amplification product in only hermaphrodite individuals. The third marker, OPA20 amplified a ~1,140-bp fragment from female and hermaphrodite DNAs, but failed to do so from the male plant DNAs.

Genetic Variation and Differences within and between Populations of Cultured and Wild Bullhead (Pseudobagrus fulvidraco) Revealed by RAPD-PCR

  • Yoon Jong-Man;Kim Gye-Woong;Park Hong-Yang
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.213-221
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    • 2005
  • We used nine decamer primers to generate DNA fragment sizes ranging from 100 bp to 1,600 bp from two bullhead (Pseudobagrus fulvidraco) populations of Dangjin in Korea. 376 fragments were identified in the cultured bullhead population, and 454 in the population of wild bullhead from Dangjin: 287 specific fragments $(76.3\%)$ in the cultured bullhead population and 207 $(45.6\%)$ in the wild bullhead population. On average, a decamer primer was used to generate 34.2 amplified products in a cultured bullhead. A RAPD primer was used to generate an average of 3.1 amplified bands per sample, ranging between 2.5 and 6.0 fragments in this population. Nine primers also generated 24 polymorphic fragments (24/376 fragment, $6.4\%$) in the cultured bullhead population, and 24 (24/454 fragments, $5.2\%$) in the wild bullhead population. The OPA-16 primer, notably, produced which 11 out of 11 bands $(100\%)$ were monomorphic in the wild bullhead population. 110 intra-population-specific fragments, with an average of 12.2 per primer, were observed in the cultured bullhead population. 99 fragments, with an average of 11.0 per primer, were identified in the wild bullhead. Especially, 55 inter-population-common fragments, with an average of 6.1 per primer, were observed in the two bullhead populations. The bandsharing value (BS value) of individuals within the wild bullhead population was substantially higher than was determined in the cultured bullhead population. The average bandsharing value was $0.596\pm0.010$ within the cultured bullhead population,. and $0.657\pm0.010$ within the wild bullhead population. The dendrogram obtained with the nine primers indicates two genetic clusters, designated cluster $1\;(CULTURED\;01\~CULTURED\;11)$, and cluster $2\;(WILD\;12\~WILD\;22)$. Ultimately, the longest genetic distance displaying significant molecular differences was determined to exist between individuals in the two bullhead populations, namely between individuals WILD no. 19 of the wild bullhead population and CULTURED no. 03 of the cultured bullhead population (genetic distance = 0.714). RAPD-PCR allowed us to detect the existence of population discrimination and genetic variation in Korean population of bullhead. This finding indicates that this method constitutes a suitable tool for DNA comparison, both within and between individuals, populations, species, and genera.

Bandsharing Values and Genetic Distances of Two Wild Shortnecked Clam, Ruditapes philippinarum Populations from the Yellow Sea Assessed by Random Amplified Polymorphic DNAs-Polymerase Chain Reaction

  • Yoon, Jong-Man;Kim, Yong-Ho
    • Journal of Aquaculture
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    • v.17 no.1
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    • pp.12-23
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    • 2004
  • Genomic DNAs were extracted from the muscle of twenty-two specimens of two shortnecked clam, Ruditapes phifippinarum populations collected in Anmyeondo and Seocheon. Genetic differences within and between populations were analysed by random amplified polymorphic DNAs-polymerase chain reaction (RAPD-PCR) using twenty arbitrary decamer primers. Out of 20 primers, 6 generated a total of 1,111 major and minor RAPD bands from individuals of two sites, producing approximately 4.2 average polymorphic bands per primer in individuals from Anmyeondo and ranging in size from less than 50 to larger than 1,500 base pairs (bp). The electrophoretic analysis of RAPD products amplified showed moderate levels of similarity among the different individuals in Seo-cheon population. The average bandsharing values (BS value) of the samples within population from Anmyeondo ranged from 0.155 to 0.684, whereas it was 0.143∼0.782 within population from Seocheon. The average BS value between individuals No. 13 and No. 14 from Seocheon was 0.782 which was higher than that of those from Anmyeondo. The single linkage dendrogram resulted from three primers (OPA-08, -09 and -20), indicating six genetic groupings composed of group 1 (No.4, 8 and 10), group 2 (No. 18), group 3 (No.2, 5 and 7), group 4 (No. 1, 3, 6, 9, 11, 12, 13, 14, 15 and 17), group 5 (16, 19 and 20) and group 6 (No. 21 and 22). In the Seocheon population, the individual No. 18 clustered distinctly from the others of this population. The observed genetic distance between the two populations from Anmyeondo and Seocheon was more than 0.209 (0.247 and 0.275). The shortest genetic distance (0.094) displaying significant molecular differences was between individuals No. 13 and No. 14. Especially, the genetic distance between individuals No. 22 and the remnants among individuals in two geographical populations was highest (0.275). This result illustrated that individual No.22 is distinct from other individuals within two shortnecked populations. The different geographical features of two sites may have caused the genetic diversity in two shortnecked clam populations.

Genetic Differences within and between Populations of Korean Catfish (S. asotus) and Bullhead (P. fulvidraco) Analysed by RAPD-PCR

  • Yoon, Jong-Man;Kim, Jong-Yeon
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.8
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    • pp.1053-1061
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    • 2004
  • Of the 20 arbitrarily chosen primers, six oligonucleotides decamer primers were used on the basis of the number of the polymorphisms generated in catfish (Silurus asotus) from Yesan and bullhead (Pseudobagrus fulvidraco) from Dangjin in Korea. Six primers were used generating a total of 602 scorable bands in catfish and 195 in bullhead population, respectively, ranging in size of DNA fragments from less than approximately 100 to larger than 2,000 base pairs (bp). Six primers yielded 199 polymorphic fragments (33.1%) in catfish and 47 (24%) in bullhead, respectively. In the present study, a total of 328 common fragments (an average of 54.7 per primer) were observed in catfish population, whereas 84 (an average of 14.0 per primer) in bullhead. The total number of specific fragments in catfish and bullhead population were 76 and 64, respectively. In catfish population, random decamer, OPA-17 (GACCGCTTGT) generated the highest number of fragments (a total of 141) in comparison with other primers used, with an average of 11.8. The common bands in the molecular weight of 300 bp generated by random primer OPA-06 (GGTCCCTGAC) were present in every individuals in bullhead population. The major polymorphic bands in the molecular weight of 100 bp generated by OPA-17 were identified in lane 14, 15, 17, 18, 19 20 and 21, which were identifying species in bullhead population. The average bandsharing values (BS values) of all of the samples within catfish population ranged from 0.575 to 0.945, whereas 0.063-1.000 within bullhead population. The bandsharing value (index of similarity between individuals) between individual No. 5 and No. 9 showed the highest level within catfish population, whereas the bandsharing value between individual No. 1 and No. 2 showed the lowest level. The single linkage cluster analysis resulted from four primers, indicating four genetic groupings composed of group 1 (C1-C10, all of the catfish samples), group 2 (B11, B12, B13, B14, B16, B17, B18, B19), group 3 (B15) and group 4 (B20 and B21). The dendrogram reveals close relationships between individual identities within two species populations and individuals derived from the same ancestor, respectively. However, genetic distances between two species populations ranged from 0.124 to 0.333. The shortest genetic distance (0.042) displaying significant molecular differences was between individual No. 6 and No. 9 catfish population. The shortest genetic distance (0.033) displaying significant molecular differences also was between individual No. 18 and No. 19 in bullhead population. Reversely, the genetic distance of individual No. 20/21 among individuals in bullhead population was highest (0.333). This result showed that bullhead No. 20 and 21 were distinct from other individuals within bullhead population.

Analysis of Genetic Diversity in Echinochloa Species Using Random Amplified Polymorphic DNAs(RAPDs) Markers (RAPD Marker를 이용한 피 수집종의 유연관계 분석)

  • Kim, Kil-Ung;Sohn, Jae-Keun;Shin, Dong-Hyun;Kim, Kyung-Min;Kim, Hak-Yoon;Lee, In-Jung
    • Korean Journal of Weed Science
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    • v.18 no.1
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    • pp.76-83
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    • 1998
  • Echinochloa species maintained by selling for more than 10 years were classified using random amplified polymorphic DNAs(RAPDs) analysis. Seventy-four decamer of randomly sequence markers were used to classify intraspecific variation irt Echinochloa species. The number of amplification products increased with increasing GC content of the primer in the range between 60% and 70% GC. Single-base substitutions of a primer altered amplification, providing new polymorphisms. The size of amplified DNA was mostly between 0.40kbp and 1.4kbp with the most common bands at 1.1kbp. Echinochloa species were detected with 6 primers which generated 26 polymorphic amplified DNAs. By hierarchical cluster analysis, Echinochloa species collected in Korea were divided into three groups. These results revealed that RAPD markers are useful tools for the determination of genetic variations in Echinochloa species.

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Genetic Differences and Variation in Two Purple Washington Clam (Saxidomus purpuratus) Populations from South and North Korea

  • Yoon, Jong-Man;Park, Su-Young
    • The Korean Journal of Malacology
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    • v.22 no.2
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    • pp.97-108
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    • 2006
  • Genomic DNA samples isolated from geographical purple Washington clam (Saxidomus purpuratus) were obtained from two different regions in Korean Peninsula: Gunsan (Gunsan population; GSP), and Haeju (Haeju population; HJP), a collection area in the vicinity of the West Sea. The seven arbitrarily primers, OPA-07, OPA-09, OPA-18, OPA-20, OPC-03, OPC-06 and OPC-09 were shown to generate the total loci, loci observed per primer, shared loci by each population, specific, and polymorphic loci which could be clearly scored. We also generated the unique shared loci to each population and shared loci by the two populations in purple Washington clam. The size of the DNA fragments also varied wildly, from 50 to 2,400 bp. Here, 304 total loci were identified in the GSP purple Washington clam population, and 282 in the HJP: 91 polymorphic loci (29.9%) in the GSP and 47 (16.7) in the HJP. 198 shared loci, with an average of 28.3 per primer, were observed in the GSP population. The decamer primer OPA-07 generated the shared loci by the two populations, approximately 1,000 bp, between the two Saxidomus populations. The oligonucleotide primer OPC-03 also generated the shared loci by the two populations, approximately 500 bp and 1,000 bp, in GSP population from Gunsan and HJP population from Haeju. The other primer, OPC-06 generated the shared loci by two Gomphina populations (approximately 400 bp). The dendrogram, generated by seven reliable primers, indicates three genetic clusters. The dendrogram obtained by the seven primers indicates three genetic clusters: cluster 1 (GUNSAN 01-GUNSAN 02), cluster 2 (GUNSAN 03-GUNSAN 11), and cluster 3 (HAEJU 12-HAEJU 22). The genetic distance between the two geographical populations ranged from 0.043 to 0.506. Especially, the longest genetic distance displaying significant molecular differences, 0.506, was found to exist between individuals GUNSAN no. 11 of Gunsan and HAEJU no. 17 of Haeju.

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Genetic Identity between Bhadawari and Murrah Breeds of Indian Buffaloes (Bubalus bubalis) Using RAPD-PCR

  • Saifi, H.W.;Bhushan, Bharat;Kumar, Sanjeev;Kumar, Pushpendra;Patra, B.N.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.603-607
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    • 2004
  • Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out with a battery of 11 random decamer primers to study band frequency (BF), genetic identity index (I) and mean average percentage difference (MAPD) between Bhadawari and Murrah breeds of buffalo. The primers OPA04 and BG15 resolved a band of 460 bp, which was present only in animals of Bhadawari breed. Whereas, the primers OPA14, BG27 and BG28 produced Murrah specific fragments of sizes 730 bp and 1,230 bp, respectively. The estimate of genetic identity index was highest (0.845) with the primer OPA01 and the lowest (0.479) with the primer BG27. The genetic identity index pooled over the primers was 0.596${\pm}$0.037 between these two breeds. The highest MAPD estimate (53.9) between the two breeds was obtained with the primer BG27 and the lowest (14.3) with the primer OPA01. It might be concluded that the genetic identity index between these two breeds calculated on the basis of BF showed moderate level of genetic identity with the primers employed. MAPD calculated on the basis of uncommon bands also demonstrated lower to medium level of genetic difference between Bhadawari and Murrah breeds of buffalo.