• Title/Summary/Keyword: random primer

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Classification of Korean Lentinula edodos Strains by Random Amplified Polymorphic DNA (RAPD) Markers (RAPD(Random Amplified Polymorphic DNA) 검정을 이용한 한국 표고균주의 계통분류)

  • Lee, Tai-Soo;Bak, Won-Chull;Kang, Ho-Duck;Kim, Se-Kwon;Byun, Byung-Ho;Yi, Chang-Keun;Lee, Won-Kyu;Min, Du-Sik
    • The Korean Journal of Mycology
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    • v.25 no.3 s.82
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    • pp.219-225
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    • 1997
  • Random Amplified Polymorphic DNA (RAPD) assay was used to identify seven typical Lentinula edodes (Berk.) Pegler strains isolated in Korea. Twenty primers from OPA-01 to OPA-20 were applied to generate the recognition of L. edodes strains. Out of 20 primers, nine primers showed efficient RAPD patterns to classify the 7 strains tested, but the rest eleven primers were not useful to be used. Even though there was no single primer that could classify all of the strains, any combination of two primers among the nine primers could identify the strains tested. Thus, RAPD assay turned out to be very precise method for classifying L. edodes strains.

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Optimum Condition of Polymerase Chain Reaction Techniques for Randomly Amplified Polymorphic DNA of Strawberry (딸기의 RAPD를 위한 PCR의 최적조건)

  • 양덕춘;최성민;강태진;이미애;송남현;민병훈
    • Korean Journal of Plant Resources
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    • v.14 no.1
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    • pp.65-70
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    • 2001
  • This study was performed to select marker which can identify genetic variation between mother plant and in vitro cultured plantlets of strawberry by PCR using random primer. When 'Yeobong' DNA extracted was treated with proteinase-K and RNase-H, clear DNA bands were shown. The optimal condition for RAPD in strawberry was to use 50ng of template DNA, 10pmol of primer,37oC of annealing temperature, and 45 cycles of PCR. After establishing above PCR optimal condition, RAPD pattern was investigated by using UBC primers. PCR was performed, and 46 of 90 primers produced PCR product showing 158 total bands. GC content was compared between the primers forming bands and no bands. The GC content showing bands was average 67.4%, whereas primers showing no bands 58%.

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A Random Amplified Polymorphic DNA (RAPD) primer to assist the Identification of Panax ginseng in Commercial Ginseng Granule Products

  • Shim, Young-Hoon;Choi, Jung-Ho;Park, Chan-Dong;Lim, Chul-Joo;Kim, Do-Hun;Cho, Jung-Hee;Kim, Hong-Jin
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.85.1-85.1
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    • 2003
  • Previously, we found the operon random primer (OP-5A) that is characteristic the genus Panax by randomly amplified polymorphic DNA (RAPD) analysis. However, OP-5A primer is limited to apply on the differentiation of only crude herbal plants. To construct more sensitive and unique primers on the genus Panax, ginseng-specific DNA profile (350 bp) that was amplified by OP-5A primer were inserted in a plasmid vector in the TA cloning method and sequenced. We designed the PCR primers (Forward: 5"-AGGGGTCTTGCTAT AGCGGAAC-3", Reverse: 5"-AGTCTTAATTTCATATTTTCGTATG-3") and identified the unique ginseng band (350 bp) in commercial granule products including ginseng extracts as well as crude ginseng plants by nascent PCR.(omitted)

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Analysis of Genetic Relationship among Cymbidium germplasms Using RAPD and URP (RAPD와 URP를 이용한 심비디움 유전자원 유연관계 분석)

  • Park, Pue Hee;Kim, Mi Seon;Lee, Young Ran;Park, Pil Man;Lee, Dong Soo;Yae, Byeong Woo
    • FLOWER RESEARCH JOURNAL
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    • v.18 no.3
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    • pp.201-206
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    • 2010
  • The genetic relationship among 48 Cymbidium cultivars was analyzed using randomly amplified polymorphic DNA (RAPD) with eighty 10 mers random primers (Operon Technologies) and twelve 20 mers random primers. Forty eight Cymbidium cultivars included 34 oriental Cymbidium, 7 hybrids, and 7 western Cymbidium. 407 (9.9 per primer) and 56 polymorphic bands (9.5 per primer) were generated by polymerase chain reaction with selected thirty 10 mers primers, and nine 20 mers primers, respectively. The polymorphic fragments ranged from 0.4 to 1.5 kb in size. The dendrogram was constructed by using the UPGMA clustering algorithm based on genetic similarity. Forty eight Cymbidium cultivars were classified into four major groups at similarity coefficient value of 0.638.

Variation of RAPD patterns between Male and Female Genomic DNAs in Dioecious Rumex acetosa L. (자웅이주 식물 수영 (Rumex acetosa L.)에서 암.수에 따른 RAPD pattern의 다양성 분석)

  • 김동순;구달회;허윤강;방재욱
    • Korean Journal of Plant Resources
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    • v.16 no.1
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    • pp.55-60
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    • 2003
  • The genetic variation of random amplified polymorphic DNA (RAPD) patterns of genomic DNAs was investigated in dioecious plant Rumex acetosa L., which carries different sex chromosome complements in female (2n=12A+XX) and male (2n=12A+XY$_1$Y$_2$). One hundred and twenty random primers consisted of 10-mer were used for PCR amplification. Polymorphic bands were found in 24 primers. Specific bands for female and male were 16 and 18, respectively. Especially, a band of 1,440 bp from the OPC-10 primer was male specific. These sex specific RAPD markers are used to understanding the sex determination mechanism in plants.

Enhanced Discrimination of Listeria spp. Using RAPD Fingerprinting Complemented by Ribotyping-PCR (리스테리아균의 특성분석을 위한 Molecular Typing 방법의 상호보완)

  • 임형근;홍종해;박경진;최원상
    • Journal of Life Science
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    • v.13 no.5
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    • pp.699-704
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    • 2003
  • The results typed by random amplification of polymorphic DNA (RAPD) were compared with those obtained by Enterobacterial repititive intergenic consensus (ERIC) fingerprinting and ribotyping-PCR. The discriminatory power of RAPD typing was the best among the methods tested. RAPD typing with two different primers for 13 Listeria spp. reference strains produced 11 patterns each. In contrast, ERIC fingerprinting produced 9 patterns and ribotyping-PCR produced 7 patterns each. Composite of two separate RAPD (Lis 11 and primer 6) results or RAPD (Lis11)/ ribotyping-PCR differentiated all 13 Listeria spp. reference strains. Therefore, composite of 2 separate RAPD (Lis11 and primer 6) or composite of RAPD (Lis11)/ribotyping-PCR is expected the most promising approach for typing field isolated Listeria spp. strains.

Studied on Amplificative Efficiency of PCR of Predigested template DNA and GC Contents for RAPD Analysis in the Silkworm, Bombyx mori (누에의 RAPD 분석을 위한 primer의 GC 함량과 사전 제한효소 처리한 주형 DNA의 PCR 증폭효율에 관한 연구)

  • 이진성;황재삼;이상몽;황석조;강현아;성승현;서동상
    • Korean journal of applied entomology
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    • v.35 no.1
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    • pp.58-65
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    • 1996
  • This experiment was carried out to evaluate the effect of sublethal doses of BPMC, etofenprox, and buprofezin on N. lugens. and its predator C. lividipennis. Buprofezin was found to be the most toxic to N. lugens and the most safe to C. lividipennis among the three insecticides, based on LD50 values. Selective toxicity index calculated by dividing LDSo value of C. lividipennis by that of N. lugens indicated that buprofezin was very safe to C. lividipennis, showing selective toxicity of 2703.3. Longevity and fecundity of N. lugens treated with LDIU and LDm of buprofezin and BPMC were not significantly different with those of untreated brown planthoppers. However, egg hatchability' of N. lugens was greatly reduced when treated with LDm of buprofezin, having the highest inhibition rate of 17.7%. Hatchability of eggs from insects treated with BPMC was similar to that of control. The oviposited peak of treated hoppers appeared late as compared to the untreated which showed the peak at early part of the ovipositional period. The longevity and fecundity of C. lividipennis treated with BPMC were significantly reduced as compared with the untreated. Etofenprox also induced fecundity reduction when treated with LDlo, and LDm. However, C. lividipennis treated with sublethal doses of buprofezin showed no redution in logevity and fecundity. From these results, it may be said that buprofezin can be used to control brown planthopper without disrupting of C. lividipennis population in the rice field.

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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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Development of Sequence Characterized Amplified Regions (SCAR) Showing for Cheju Native Horse (품종 특이성을 이용한 제주마 판별 표지인자 재발)

  • Cho Byung Wook
    • Journal of Life Science
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    • v.15 no.3 s.70
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    • pp.474-478
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    • 2005
  • This study was conducted to analyze genetic characteristics and to develop the specific marker for Cheju native horse (Coo) at the level of sequence characterized amplified regions (SCARs). We collected blood samples from Cheju native horse and Thoroughbred horse (Th) and obtained genomic DNA from the blood of 50 individuals randomly selected within the breeds. Seven hundred primers were chosen randomly and were used to examin the polymorphism and 40 kinds of primers showed polymorphic RAPD band patterns between two breeds. Thirty primers of them showed horse specific bands. With the primer MG 30, amplified band of 2.0 kb showed the specificity to Cheju native horse (Cnh). Additionally MG 53 detected the thoroughbred horse (Th) specific markers at size of 2.3 kb. As the next, 2.3 kb band from MG 53 was checked with the all individuals from all the breeds of this study, and it maintained the reproducible breed specificity to thoroughbred horse (Th). With this results, 2.3 kb band was cloned into plasmid vector and sequenced bidirectionally from both ends of the cloned fragment. With the obtained sequences 10 nucleotide extended primers including the original arbitray primer were designed as a SCARs primer. Finally, the primer with extended sequence showed the reproducible breed differentiation pattern and it was possible to identify Cheju native horse (Cnh) from other breeds. The SCARs marker 2.3 kb from MG 53 could be used to identify Cheju native horse (Cnh) for not only registration but also horse breeding programe.

Analysis of Genetic Relationships of Korean Native Polygonatum spp. (우리 나라 자생 둥굴레속 식물의 유연관계 분석)

  • Jang, Kye-Hyun;Song, Geun-Woo;Chung, Jong-Il;Kang, Jin-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.12 no.3
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    • pp.214-218
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    • 2004
  • Germplasms having valuable characters are increasjngly important for modern breeding programs. This study was conducted to obtain the basic data for effective use of genetic resources of Polygonatum. The relationship of seven Polygonatum species collected widely in Korea was analyzed by RAPD markers. Total number of alleles amplified by nineteen random primers were 114 to 157, and variation in number of alleles was also diverse among seven species examined. The seven species were divided into two groups; one was of Polygonatum stenophyllum and Polygonatum humile. the other was of Polygonatum inflatum, Polygonatum lasianthum, Polygonatum odoratum var. pluriflorum (1), (2), and for variegatum.