• Title/Summary/Keyword: RAPD primer

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Identification of DNA polymorphisms in the field bean ( Glycine soza S. and Z. ) using RAPD markers (RAPD 표지인자를 이용한 돌콩 DNA 다형현상 분석)

  • 이성규
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.2
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    • pp.143-150
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    • 1998
  • Six field bean (GI-vcine soza S and Z ) plants were examined for their genetic polymorphisms and intraspecific variations using randomly amplified polymorphic DNA(RAPD) markers. In RAPD analysis of 5 random primers (Rp-1, Rp2, Rp-3, Rp-4, Rp-5), 30 of total 155 bands obtained kom 5 primers were polymorphic and sizes of polymirphic band ranged between 0.5 and 3.0 kb. Number of bands amplyfied per primer was varied from 2 to 11 and average number was 6.0. Genetic variation of intraspecies in the samples of six region was ranged behveen 11 to 25 percent, and genetic similarity among intraspecies was ranged from 0.69 to 0.78. In pairwise genetic similarity test of six field bean plants, Mun and Hoj showed highest coefficient of genetic similarity as 0.67, whereas Sin and Hoj was lowest as 0.45. According to the genetic similarity, the level of intraspecific variation is higher than that of regional distance in GI-vcine soza.

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Interspecific relationships of Korean Viola based on RAPD, ISSR and PCR-RFLP analyses (RAPD, ISSR과 PCR-RFLP를 이용한 한국산 제비꽃속(Viola)의 종간 유연관계)

  • Yoo, Ki-Oug;Lee, Woo-Tchul;Kwon, Oh-Keun
    • Korean Journal of Plant Taxonomy
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    • v.34 no.1
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    • pp.43-61
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    • 2004
  • Molecular taxonomic studies were conducted to evaluate interspecific relationships in Korean Viola 34 taxa including two Japanese populations using RAPD(randornly amplified polymorphic DNA), ISSR(inter simple sequence repeat) and PCR-RFLP(restriction fragment length polymorphism) analysis. Only six and four primers out of 40 arbitrary and 12 ISSR primers were screened for 34 taxa, and were revealed 70 (98.6%) and 28 (96.6%) polymorphic bands, respectively. Fifteen restriction endonucleases produced 80 restriction sites and size variations from the large single copy region of cpDNA, 16 (20%) of which were polymorphic. The separate analyses from the RAPD, ISSR and PCR-RFLP data were incongruent in the relationships among 34 taxa, but combined data was in accordance with previous infrageneric classification system based on morphological characters, especially the subsection and series level. Section Chamaemelanium placed between subsect. Patellares and Vagimtae of section Nomimium was not formed as a distinct group. Viola alb ida complex including three very closely related taxa was recognized independent group within subsect. Patellares in combined data tree. This result strongly suggested that they should be treated to series Pinmtae. RAPD analysis was very useful to clarify the interspecific relationships among the species of Korean Viola than ISSH and PCR-RFLP analyses.

A Study of Optium Condition of RAPD for the Analysis of Genetic Characteristics by Autumn Leaf Color of Zelkova serrata (느티나무(Zelkova serrata)단풍의 유전적 특성분석을 위한 RAPD 적정 조건 구명에 관한 연구)

  • Choi, Byoung Kon;Bang, Kwang Ja
    • Journal of the Korean Society of Environmental Restoration Technology
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    • v.7 no.5
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    • pp.94-99
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    • 2004
  • This study was carried out to find out what is the optimum conditions for RAPD of Zelkova serata. We changes the factors what affect to PCR band patterns, as a result, we established the optimum conditions as follows; template DNA 100mg, Primer 0.25uM, dNTP 100mM, Taq polymerase 1.0u, and total reaction volume was filled up to 10uL with distilled water. As the amount of primers went higher, PCR reaction rates were lowered. This reason was cause by exhaustion of primers during initial reaction. The amount of dNTP didn't showed noticable differtations between the range, but the optimum amount was 100mM for efficiency. Taq polymerase 1.0 unit was the best in the range. As the concentration of polymerase were increased, many non-specific bands were appeared, In primer selection, most Openron Random Primers are amplified in this experiment. The primers GC contents were 60, and set A, B, C, D, E, X were tested. Thermal cycler(ASTEC PC808, Japan) condition was, $95^{\circ}C$, 5min, initial denaturation, $94^{\circ}C$, 20sec, denaturation, $37^{\circ}C$, 40sec, annealing, $72^{\circ}C$, 1min, extention, 45cycle repeated and final extention $72^{\circ}C$10min.

Intraspecific Relationship of Eleutherococcus senticosus Max. by RAPD Markers (가시오갈피 수집종의 RAPD 변이분석)

  • Kim, Sun;Kim, Ki-Young;Park, Mun-Su;Choi, Sun-Young;Yun, Song-Joong
    • Korean Journal of Medicinal Crop Science
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    • v.6 no.3
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    • pp.165-169
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    • 1998
  • To evaluate the intraspecific variations among the Kasiogalpi(Eleutherococcus senticosus Max.) collections. randomly amplified DNA polymorphisms were examined. Twenty primers from 90 primers applied were selected. The range of polymorphism was $7.1{\sim}90.9%$ in 113 randomly and specifically amplified DNA fragments. Collections were divided into two major groups at the similarity coefficient value of 0.65. A considerable degree of genetic diversity was also detected among plants within the same collections. Deokyu (1, 2, 3, 4, 6), Bukhaedo (7, 8) and Odae (9, 10) col1ections showed higher degree of genetic similarity with a value of $0.65{\sim}0.86$, while Deokyu 5 showed much lower genetic similarity than other col1ections.

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Growth and DNA Alteration of Heavy-ion Beam Irradiated Tobacco(Nicotiana plumbaginifolia) Plant (중이온 빔조사 담배(Nicotiana plumbaginifolia) 식물체의 생장과 DNA 변이)

  • Lyu Jae-Il;Kim Min-Su;Tomoko Abe;Lee Hyo-Yeon;Yang Deok-Chun;Bae Chang-Hyu
    • Korean Journal of Plant Resources
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    • v.18 no.1
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    • pp.169-178
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    • 2005
  • Effects of heavy-ion beam$(^{20}Ne)$ irradiation on growth and DNA alteration of tobacco plants were investigated. Seed germination and plant height were decresed as the ion-beam intensity was increased. However, the bolting and flowering were promoted by the low intensities of 5 Gy to 10 Gy treatment. Out of the 100 primers screened, 59 primers generated 336 DNA fragments by RAPD analysis, and one specific DNA fragment that amplified in control but not in the ion-beam irradiated plants was observed. By AFLP analysis, DNA fragment difference related to the ion-beam treatment was not detected but observed among the plant bodys.

Intraspecific Relationship Analysis of Eleutherococcus senticosus Max. by RAPD Markers (RAPD 분석에 의한 가시오갈피의 유연관계 분석)

  • 임정대;성은수;최강준;김승경;김명조;유창연
    • Korean Journal of Plant Resources
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    • v.13 no.2
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    • pp.104-110
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    • 2000
  • To analyse the genetic relationship and intraspecific variations among the Eleutherococcus senticosus population, the polymerase chain reaction(PCR) was performed total genomic DNAs of 10 E. senticosus collections by random 10 primers. The genetic diversity and genetic distance among 10 collections of Eleutherococcus spp. were used to describe the dendrogram showing phylogenic relationship. Ten collections were classfied into two group(group I, II) at the similarity coefficient value of 0.50. Group I included E. senticosus of Bukhado(Japanese), youngwal(Korea), E. seoulense, and E. chiisanesis while group II included several internal and Russia collection. The range of polymorphism was from 66.7 to 90.9% in 87 amplified DNA fragments. The similarity value of all collections ranged from 0.41 to 0.92. The average of genetic distance was 0.61.

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RAPD Fingerprinting for the Species Identification of Animals

  • Huang, Mu-Chiou;Horng, Yan-Ming;Huang, Hsiu-Lin;Sin, Yen-Long;Chen, Ming-Jaw
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1406-1410
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    • 2003
  • The studies were based on the RAPD fingerprinting for the species identification of animals. The genomic DNA samples of ostriches, Taiwan local chickens, Aboracres broilers, Leghorn chickens, quails, doves, emus, Beltville small white turkeys, pheasants, Chinese geese, mule ducks, Holstein cattle and Landrace pigs were amplified with random primers by RAPD-PCR for fingerprinting. The results showed that the varied band patterns of DNA fingerprints were generated from templates depending on the kinds of primers or animal species. The same primer applied to the same breed, all of the main bands are similar, but which were different among species. In order to try to identify the species from the mixture of meat by RAPD fingerprinting, the meat of ostrich and cattle was mixed in different ratios for this study. The results showed that it could be easily and precisely distinguished according to the band distribution of RAPD patterns.

Analysis of Genetic Relationships in Hypericum erectum Thunb. by RAPD (RAPD 방법을 이용한 고추나물의 유연관계분석)

  • Kim, Sun-Hee;Kim, Eung-Sik;Kim, Sung-Ho;Ahn, Jun-Cheul;Hwang, Baik
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.4
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    • pp.141-145
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    • 2005
  • The genetic relationships of Hypericum erectum, H. ascyron and H. perforatum Thunb. by RAPD using total 46 primers were analysed 30 primers among primers tested showed the amplification band in all. The amplified DNA fragment ranged from 0.25 to 6.6 kb in size. The 411 bands (34.4%) among 1194 bands derived from 30 primers were polymorphic, and 13.7 bands per primer were observed. The phenograms for six analyzed individuals by RAPD markers were not matched well with those of the result by morphological characters. They were clustered monophyletic at the similarity coefficient value ranged from 0.24 to 0.96.

Identification of Genetic Markers for Korean Native Cattle (Hanwoo) by RAPD Analysis

  • Yeo Jung Sou;Lee Ji Sun;Lee Chang Hee;Jung Young Ja;Nam Doo Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.23-26
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    • 2000
  • In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

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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.