• Title/Summary/Keyword: random primer

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Discrimination of Phlomidis Radix and Dipsaci Radix using the Random Amplified Polymorphic DNA Analysis (Random Amplified Polymorphic DNA 분석을 이용한 한속단과 천속단의 감별)

  • Lee, Mi-Young;Ryuk, Jin-Ah;Kim, Hong-Jun;Kim, Young-Hwa;Chae, Byoung-Chan;Ko, Byoung-Seob
    • Korean Journal of Oriental Medicine
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    • v.13 no.1 s.19
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    • pp.147-152
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    • 2007
  • As a result to amplifying 12 samples of 'Sok-dan' through an random amplified polymorphic DNA (RAPD) method using eighteen DEC and URP primers, distinct band forms enabling discrimination of Phlomus umbrosa and Dipsacus asperoides were observable in the UBC 320 primer, UBC 367 primer, UBC 385 primer, UBC 414 primer, UBC 423 primer, URP 3 primer, URP 5 primer and URP 9 primer. The polymorph result amplified with a random primer was evaluated through Gelcompar II, showing a result dividable into two groups. The divided groups were the dried sample group of Dipsacus asperoides and the group of Phlomis umbrosa. In order to recognize the distinction between Dipsaci Radix types, the genetic variation of 'Sok-dan' produced domestically and imported was evaluated through RAPD, and the potential to distinguish these in forms of dried medicine was identified, presenting a method to authentification of Phlomis umbrosa and Dispacus asperoides.

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Primers for typing Listeria spp. using Random Amplified Polymorphic DNA (RAPD) ANalysis (Listeria spp.의 RAPD typing을 위한 Primer의 분리력 비교)

  • 임형근;홍종해;박경진;최원상
    • Journal of Food Hygiene and Safety
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    • v.18 no.2
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    • pp.67-72
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    • 2003
  • Random amplified Polymorphic DNA (RAPD) analysis Is based on the amplification of random DNA segment using a single arbitrary primer. Polymorphic DNA patterns identified by this method can be used for typing Listeria monocytogenes. To select the primers for RAPD typing Listeria spp., the performance of 31 primers were compared by analyzing 13 Listeria spp. reference strains. Reproducible electrophoresis patterns were obtained. Among 31 primers, 6 primers (primer 6, HLWL74, UBC155, UBC127, Lis5, Lis11) showed better differentiation, when discrimination index, band clarity, band number, difficulty of band scoring were considered than the others. These primers will be useful far typing Listeria spp. in the future. Currently, we are under investigation for the RAPD typing of contaminated L. monocytogenes for the risk analysis of pork processing plant using these primers.

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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RAPD marker를 이용한 참돔 집단의 유전적 특성 분석

  • 장요순;노충환;홍경표;명정구;김종만
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.34-34
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    • 2003
  • 한국산 선발계통 및 일본산 양식계통과 이들 두 계통간 잡종 참돔 집단의 유전적 특성을 분석하기 위하여, RAPD (Random Amplified Polymorphic DNA) marker를 탐색하였다. 10개의 염기로 이루어진 200개의 random primer 분석을 통하여 polymorphic pattern을 나타내는 23개의 random primer를 선발하였으며, 각 primer의 재현성을 확인하였다. 이들 중 OPA-11 primer는 크기가 각각 600 bp, 650 bp 및 750 bp 인 3개의 DNA 단편에 의하여 4개의 genotype을 나타냈으며, 각 genotype의 빈도는 집단간차이를 보였고, 한국산 선발계통 집단에서는 4개의 genotype이 모두 발견되는 반면, 일본산 양식계통 및 일본산 양식계통을 포함한 교배집단에서는 특정 genotype만 발견되었다. OPA-11 primer 유래의 polymorphic DNA 단편을 cloning하고 염기서열을 결정하였으며, SCAR (Sequence Characterized Amplified Region) primer를 제작하고 분석하였다. 본 연구는 참돔집단의 유전적 특성 파악 및 집단 구별에 RAPD marker를 활용하였으며, 참돔 육종시 형질 및 기능관련 DNA marker 탐색에 적용하기 위하여, 이후의 연구에서는 SCAR과 RFLP 분석에 RAPD marker를 이용하여 100% 정확도를 갖는 RFLP maker를 찾고, MAS (Marker-Assisted Selection)에 적용하고자 한다.

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Comparison of RAPD Profiles and Phenotypical Characters of Streptococcal Strains (연쇄상구균의 표현형적 특성과 RAPD profiles 비교)

  • Song, Jin-Gyeong;Kim, Jong-Hun;Kim, Eun-Hui
    • Journal of fish pathology
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    • v.16 no.1
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    • pp.51-59
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    • 2003
  • Streptococcal infection is one of the most serious disease of cultured olive flounder, Paralychthys olivaceus in Korea and caused by more than one species. However, there has been considerable confusions about the taxonomic position of the fish pathogenic streptococci. In this study, We performed the randomly amplified polymorphic DNA(RAPD) pattern analysis to evaluate the possible classification in 8 streptococci isolated from diseased olive flounder and reference strains based on their DNA structure. RAPD PCR with DNA solution prepared by simple boiling and 10-mer random primer was appeared to be a good tool for discrimination of different streptococcal strains. Phenotypical characters by simple biological test and API 20 Strep corresponded well to the specific profiles of RAPD in streptococcal isolates of this study. Therefore, the RAPD profile was considered as one of differential characters to discriminate the streptococcal isolates from diseased olive flounder.

Primers for Typing Salmonella spp. using Random Amplified Polymorphic DNA (RAPD) Analysis (Salmonella spp.의 RAPD Typing을 위한 Primer의 분리력 비교)

  • Lim, Hyung-Kum;Lee, Kyung-Hee;Hong, Chong-Hae;Park, Gyung-Jin;Choi, Weon-Sang
    • Journal of Food Hygiene and Safety
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    • v.18 no.4
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    • pp.224-228
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis is based on the amplification of random DNA segment using a single arbitratrary primer. For typing Salmonella spp., polymorphic DNA patterns identified by this method can be used. To select the primers for RAPD typing Salmonella spp., the performances of 20 primers were compared by analyzing 16 Salmonella spp. reference strains. Reproducible electrophoresis patterns were obtained. Among the 20 primers tested, 4 primers (A, OPG04, OPG10, OPL03) showed better differentiation than the others. At the time discrimination index, band clarity, band number and difficulty of band scoring were considered. These primers will be useful for typing Salmonella spp. in the future. Curretly, we are under investigation for the RAPD typing of contaminated Slmonella spp. for the risk analysis of pork processing plant using the primers.

Identification of Korean Native Goat Meat using DNA Analysis (DNA분석기법을 이용한 한국재래산양육의 판별)

  • Sang, B.C.;Lee, S.H.;Ryoo, S.H.;Seo, K.W.;Han, S.W.;Kim, S.K.
    • Korean Journal of Agricultural Science
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    • v.26 no.2
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    • pp.33-38
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    • 1999
  • This study was carried out to analyze the genetic polymorphisms of genomic DNA of blood and meat for conservation of the genetic resources and genetic improvement of Korean Native goat. The genetic identification between Korean Native goat and imported goat was examined using RAPD(random amplified polymorphisms DNAs) analysis with 30 Korean Native goat, 10 hybrid, 10 imported goat. 10 Korean native goat meat and 10 imported goat meat. The results obtained from this study were summarized as follows: 1. Genomic DNA from Korean native goat, hybrid and imported goat could be obtained above about 23kb size using 0.5% agarose gel electrophoresis and the ratio of optical density at 260nm to that at 280nm was between 1.7 and 2.0 using UV spectrophtometer instrument. 2. In the results of the gene identification between Korean Native goat and hybrid, and imported goat using RAPD methods with random primer of 110 kinds, only Korean native goat showed a specific band at about 369bp using a random primer OPO-19 (5'-CAA ACG TCG G-3'), but imported goat and hybrid not showed. 3. Also, in the results of the gene identification between Korean Native goat meat and imported goat meat using RAPD methods with random primer, Korean native goat only showed a specific band at about 369bp using a random primer No. 19(5'-CAA ACG TCG G-3'), but imported goat not showed.

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Identification of Monoecious and Dioecious Plants of. Schisandra nigra Using the RAPD Markers (RAPD 표지인자를 이용한 흑오미자의 자웅동주 및 자웅이주 식물의 동정)

  • 이효연;한효심;이갑연;한상섭;정재성
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.309-313
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    • 1998
  • RAPD (Random Amplified Polymorphic DNA) analysis was conducted to Schisandra nigra plants in order to select the specific markers for monoecious and dioecious individuals. RAPD results using eighty random 10-mer primers revealed that S. nigra had a different banding pattern from S. chinensis and Kadsura japonica. When DNA isolated from leaves of monoecious and dioecious plants were used as PCR template, only five primers, OPA-17, OPA-19, OPB-03, OPB-09 and OFB-16, showed polymorphic band patterns. No variation in banding profiles within male or female individuals was observed when these five primers were used whereas three monoecious plants (No 1, No 2 and No 3) showed different banding patterns one another, A 750 bp segment was amplified by primer OPB-3 from male individuals. On the other hand, two segments, 950 bp and 1690 bp, with OPA-19 and 700 bp of segment with OPB-3 were amplified in female individuals. These result indicate that the specific buds of male and female S. nigra could be used as genetic markers for the early discrimination of male and female individuals.

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Genetic Diversity Analysis of the Cheju Horse Using Random Amplified Polymorphic DNAs (PCR-RAPD를 이용한 제주말의 유전적 다양성분석)

  • Cho, Byung-Wook;Lee, Kil-Wang
    • Journal of Life Science
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    • v.14 no.3
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    • pp.521-524
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    • 2004
  • This experiment was carried out to analyze genetic characteristics and to develop the breed specific DNA marker for Cheju-native horse. If this marker contains high repetitive sequences, it is possible to convert a RAPD marker of interest into a single-locus PCR marker called a sequence characterized amplified region(SCAR). Twenty six Cheju-native horse and Fifty thoroughbred genomic DNA were pooled and PCR. were accomplished using 800 random primers. Comparing the pooled DNA from Cheju-native horse and thoroughbred, we found 9 primers which identified markers present in the pooled DNA from breed but absent in the other breed. Among 9 random primers, 6 primers were thoroughbred specific and 3 primers were Cheju-native horse specific. Testing individual horse revealed that 5 marker showed the similar band pattern between Cheju-native horse and Thoroughbred. However, 4 marker were wholly absent in breed while present in the other breed. UBC $126_{3500bp}$, UBC $162_{500bp}$, and UBC $244_{1200bp}$ was detected only Thoroughbred and UBC $562_{560bp}$was detected Cheju-native horse, respectively. After determining of the cloned breed-specific fragment sequence, we designed the SCAR-primers and carried out PCR. Compared to random primer, RAPD-SCAR primer didn't show significantly higher specific band. However, RAPD analysis is useful for genetic characterization of Cheju-native horse.

Genetic Similarity and Difference between Common Carp and Israeli Carp (Cyprinus carpio) Based on Random Amplified Polymorphic DNAs Analyses

  • Yoon, Jong-Man
    • Animal cells and systems
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    • v.5 no.4
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    • pp.333-339
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    • 2001
  • Common carp (Cyprinus carpio) and its aquaculture breed Israeli carp samples were obtained from two separate aquaculture facilities under the similar raising conditions during two years in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp for identification of genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA using arbitrary primers. The arbitrary primer No.21 (ACTTCGCCAC) yielded the highest number of fragments with the average of 15.0 among the primers used in Israeli carp. A tota1 of 294 polymorphic products in common carp and 336 in Israeli carp were observed by random primers. The average number of polymorphic products generated by random RAPD primer No. 2 (GTAGAC-CCGT) showed 8.0 in Israeli carp. On average, each random RAPD primer produced 5.4 amplified polymorphic products in common carp and 6.2 in Israeli carp. An average genetic similarity (BS value) was 0.44$\pm$0.05 within the common carp and 0.32$\pm$0.04 within the Israeli carp. The degree of similarity frequency (BS) between two carps was 0.67 as generated by the primer No. 19 (GACGGATCAG). The average level of bandsharing was 0.57$\pm$0.03 between the two carps. Accordingly, the two carp populations were genetically a little distant. The electrophoretic analysis of PCR-RAPD products showed middle levels of variation between the two carp populations. This result implies that the genetic diversity among intra-population may be higher when compared with that between the two carps. The RAPD polymorphism generated by these random primers might be used as a genetic marker for populations or lines identification in important aquacultural carp.

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