• Title/Summary/Keyword: random amplification of polymorphic DNA (RAPD)

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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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Application of RAPD markers for characterization of ${\gamma}$-ray-induced rose mutants and assessment of genetic diversity

  • Chakrabarty, D.;Datta, S.K.
    • Plant Biotechnology Reports
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    • v.4 no.3
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    • pp.237-242
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    • 2010
  • Six parent and their 12 gamma ray-induced somatic flower colour mutants of garden rose were characterized to discriminate the mutants from their respective parents and understanding the genetic diversity using Random amplification of polymorphic DNA (RAPD) markers. Out of 20 primers screened, 14 primers yielded completely identical fragments patterns. The other 7 primers gave highly polymorphic banding patterns among the radiomutants. All the cultivars were identified by using only 7 primers. Moreover, individual mutants were also distinguished by unique RAPD marker bands. Based on the presence or absence of the 48 polymorphic bands, the genetic variations within and among the 18 cultivars were measured. Genetic distance between all 18 cultivars varied from 0.40 to 0.91, as revealed by Jaccard's coefficient matrix. A dendrogram was constructed based on the similarity matrix using the Neighbor Joining Tree method showed three main clusters. The present RAPD analysis can be used not only for estimating genetic diversity present in gamma ray-induced mutants but also for correct identification of mutant/new varieties for their legal protection under plant variety rights.

Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes (RAPD-PCR(Random Amplified Polymorphic DNA - Polymerase Chain Reaction) 방법을 이용한 Listeria monocytogenes의 검색)

  • Park Bum-Joon;Sihn Eon-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.17 no.3
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    • pp.254-259
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    • 2004
  • Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.

RAPD Analysis for Genetic Diversity of Melon Species (참외와 멜론의 유전적 다양성에 대한 RAPD 분석)

  • Mo, Suk-Youn;Im, Sung-Hee;Go, Gwan-DaI;Ann, Chong-Mun;Kim, Doo Hwan
    • Horticultural Science & Technology
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    • v.16 no.1
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    • pp.21-24
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    • 1998
  • RAPD markers were analyzed in order to detect the genetic variation and diversity of the fifty-two melon lines. SDS extraction method produced more and purer DNA than CTAB method. RAPD reaction conditions were optimized as follows ; 10ng template DNA, 270nM primer, $200{\mu}M$ each of dATP, dCTP, dGTP and dTTP, $0.3{\mu}unit$ dynazyme and 10x buffer brought to $15{\mu}l$ final volume with distilled water. The adequate annealing temperature was $39^{\circ}C$ and forty cycles of amplification produced the best RAPD band patterns. Among a total of 123 bands from 12 random primers, 25 polymorphic bands(20%) were selected as reliable markers. The average number of polymorphic bands per primer was 2.1 among the 52 lines. Intragroup genetic relationship based on the marker difference was closer than intergroup genetic relationship. The 52 lines could be grouped into two major group (Korean landraces and melon lines) and then melon group subdivided into two subgroups (net melon lines and no-net melon). This result corresponded to morphological grouping. Eight RAPD markers separated the Korean landraces and melon groups and four RAPD markers separated net melon and no-net melon groups.

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Mycelial Propagation and Molecular Phylogenetic Relationships of Commercially Cultivated Agrocybe cylindracea based on ITS Sequences and RAPD

  • Alam, Nuhu;Kim, Jeong-Hwa;Shim, Mi-Ja;Lee, U-Youn;Lee, Tae-Soo
    • Mycobiology
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    • v.38 no.2
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    • pp.89-96
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    • 2010
  • This study evaluated the optimal vegetative growth conditions and molecular phylogenetic relationships of eleven strains of Agrocybe cylindracea collected from different ecological regions of Korea, China and Taiwan. The optimal temperature and pH for mycelial growth were observed at $25^{\circ}C$ and 6. Potato dextrose agar and Hennerberg were the favorable media for vegetative growth, whereas glucose tryptone was unfavorable. Dextrin, maltose, and fructose were the most effective carbon sources. The most suitable nitrogen sources were arginine and glycine, whereas methionine, alanine, histidine, and urea were least effective for the mycelial propagation of A. cylindracea. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The sequence of ITS2 was more variable than that of ITS1, while the 5.8S sequences were identical. The reciprocal homologies of the ITS sequences ranged from 98 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) using 20 arbitrary primers. Fifteen primers efficiently amplified the genomic DNA. The average number of polymorphic bands observed per primer was 3.8. The numbers of amplified bands varied based on the primers and strains, with polymorphic fragments ranging from 0.1 to 2.9 kb. The results of RAPD analysis were similar to the ITS region sequences. The results revealed that RAPD and ITS techniques were well suited for detecting the genetic diversity of all A. cylindracea strains tested.

Absence of DNA Polymorphisms in Myzus persicae (Homoptera: Aphididae) in Relation to their Host Plants (기주식물 종류에 따른 복숭아혹진딧물(Myzus persicae)의 DNA Polymorphism 비교)

  • H. J. Kim;K. S. Boo;K. H. Cho
    • Korean journal of applied entomology
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    • v.35 no.3
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    • pp.209-215
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    • 1996
  • DNA polymorphisms were analyzed for 8 clones of the green peach aphid, Myzus persicae Sulzer, by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR). The insect has different host preferences and was even classified into two different species, M. persicae Sulzer and Myzus nicotinae Blackman by their morphological characters, but this point is still in arguement. To identify the differences between two types of the green peach aphid by RAPD-PCR, the template DNA was extracted from 4 clones each of tobacco-feeding and non-tobacco-feeding forms and one hundred primers of 10-nucleotideslong were tested in PCR. The amplified DNAs were analyzed by agarose gel electrophoresis. Eighty-three primers gave amplified DNA fragments with 1 to 22 in number and 500 to 20,000 base pairs in length, but no amplification was observed in the other 17 primers. The average number of fragment per each amplification was about 13. In the case of 82 out of 83 random primers, band patterns of amplified DNA were identical among 8 clones, even though some differences were noticed in the intensity of specific bands. Polymorphism was detected by only one primer within the tobacco-feeding forms, but not between the two host types. The results did not detect any relationship between RAPD polymorphism and their host preference.

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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.

Genetic Similarity Frequency and DNA Polymorphism between Common Carp and Israeli Carp Using Polymerase Chain Reaction-Random Amplified Polymorphic DNAs

  • Yoon, Jong-Man;Park, Min-Soon;Kim, Young-Gill
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.334-335
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    • 2001
  • Common carp (Cyprinus carpio) and Israeli carp(C. carpio) samples were obtained from a aquaculture facility in the Kunsan National University, Korea. Genomic DNA was isolated from the common carp and Israeli carp representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. There were observed a total of 90 species-specific genetic markers within Israeli carp. On average, each random RAPD primer produced amplified 7.9 products from 1 to 17 bands. An average genetic similarity within Israeli carp showed -.60$\pm$0.05. The average level of bandsharing was some 0.57$\pm$0.03 between common carp and Israeli carp. Accordingly, two carp species were genetically a little distant. The electrophoretic analysis of PCR-RAPD proudcts showed high levels of variation between two fish species. The RAPD polymorphism generated by primer may be used as a genetic marker for species or lines identification in important aquacultural carp.

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Analysis of Genetic Polymorphisms and Similarity Using Random Amplified Polymorphic DNAs in Cattle (RAPD기법을 이용한 축우의 유전적 다형성과 유사도 분석)

  • Lee, S.H.;Seo, K.W.;Kwon, I.;Sung, C.K.;Kim, S.K.;Sang, B.C.
    • Korean Journal of Agricultural Science
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    • v.26 no.2
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    • pp.39-48
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    • 1999
  • This study was carried out to investigate the difference and genetic similarity at the level of molecular genetics. Genomic DNA was extracted from blood of Holstein, Korean cattle, Charolais, and hybrid between Korean cattle and charolais and RAPD(random amplified polymorphic DNAs) was analyzed by PCR(polymerase chain reaction). After genetic similarity value from different breeds are analyzed, genetic similarity was estimated by UPGMA(unweighted pair-group method using average). The results obtained from this study can be summarized as follows: 1. When genomic DNA which was extracted from different breeds was subjected to electrophoresis on 1.5% agarose gel, bigger than 12.2kb was appeared. Ratio by absorbance of $A_{260}/A_{280}$ was 1.75~2.10, indicating that genomic DNA was quite pure for RAPD analysis. 2. Different band patterns by RAPD were appeared according to the breeds in cattle. The best primer used to distinguish Holstein from other breeds was 5'-GAC CGC TTG T-3'. 3. A 340bp fragment was amplified in $33.0^{\circ}C$ of annealing temperature for the Holstein and Charolais breeds, but any amplification was not occurred in this annealing temperature for Korean cattle and hybrid. In addition, a 340bp fragment was amplified in $37.5^{\circ}C$ of annealing temperature for the Holstein and Korean cattle, but any amplification was not occurred in this annealing temperature for Charolais and hybrid. For the reaction of PCR. $37.5^{\circ}C$ and $33.0^{\circ}C$ of annealing temperature was shown to be best for genetic marker identification from Holstein, Charolais, and hybrid between Korean cattle and Charolais. 4. When genetic similarity from different breeds are analyzed at the both temperature of $33.0^{\circ}C$ and $37.5^{\circ}C$, the genetic similarity value of Holstein and Korean cattle, Holstein and Charolais, Korean cattle and Charolais, and Korean cattle and hybrid were 0.666~0.777, 0.615~0.666, 0.400~0.461 and 0.857~0.888, respectively. 5. It could be concluded that different breeds are capable of distinguishing by RAPD used random primer 5'-GAC CGC TTG T-3', genetic similarity from different breeds was appeared the higher genetic similarity value of Korean cattle and Charolais than that of Holstein between Korean cattle and Charolais by UPGMA.

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Seed Purity Test and Genetic Diversity Evaluation Using RAPD Markers in Radish (Raphanus sativus L.)

  • Huh, Man-Kyu;Choi, Joo-Soo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.4
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    • pp.346-350
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    • 2009
  • The cultivated radish (Raphanus sativus L.) is a major vegetable crop in the world wide and fast-growing species that grows inhabitats of six continents. It is very important to determine hybrid seed purity in the production of hybrid Brassica vegetable seeds to avoid unacceptable contamination with self-inbred (sib) seeds. The use of random amplified polymorphic DNA (RAPD) markers for evaluating seed purity in $F_2$-hybrid radish cultivars demonstrated. One hundred eighty seeds from the F1 male and female harvest were subsequently screened for seed purity using 13 primers. The 13 primers result in 17 cultivar-specific bands and 23 variable RAPD bands scored for cultivar. RAPD analysis of hybrid seeds from the harvest revealed 128 seeds tested except underdevelopment and decayed seeds were sibs. Especially, $F_2$ hybrids of radish, OPC13, OPD20 were presented clear hybrid bands. It maintains higher than average level of genetic diversity compared with their correspondent parents. RAPD amplification of DNA extracted from germinated individuals from the female harvest reveal that 10 of 208 seeds tested were self-inbred (4.8%). RAPD analysis of hybrid seeds from the male harvest revealed 7 of the 208 seeds tested were sibs (3.4%). The RAPD may lead to a better insight in to the hybrid seed purity.