• Title/Summary/Keyword: RAPD Marker

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A Genetic Linkage Map of Soybean with RFLP, RAPD, SSR and Morphological Markers

  • Kim, Hong-Sik;Lee, Suk-Ha;Lee, Yeong-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.45 no.2
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    • pp.123-127
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    • 2000
  • The objective of this study was to develop a linkage map of soybean under the genetic background of Korean soybean. A set of 89 F/sub 5/ lines was developed from a cross between 'Pureunkong', which was released for soy-bean sprout, and 'Jinpumkong 2', which had no beany taste in seed due to lack of lipoxygenase 1, 2, and 3. A linkage map was constructed for this population with a set of 113 genetic markers including 7 restriction fragment length polymorphism (RFLP) markers, 79 randomly amplified polymorphic DNA (RAPD) markers, 24 simple sequence repeat(SSR) markers, and 3 morphological markers. The map defined approximately 807.4 cM of the soybean genome comprising 25 linkage groups with 98 polymorphic markers. Fifteen markers remained unlinked. Seventeen linkage groups identified here could be assigned to the respective 13 linkage groups in the USDA soybean genetic map. RFLP and SSR markers segregated at only single genetic loci. Fourteen of the 25 linkage groups contained at least one SSR marker locus. Map positions of most of the SSR loci and their linkages with RFLP markers were consistent with previous reports of the USDA soybean linkage groups. For RAPD, banding patterns of 13 decamer primers showed independent segregations at two or more marker loci for each primer. Only the segregation at op Y07 locus was expressed with codominant manner among all RAPD loci. As the soybean genetic map in our study is more updated, molecular approaches of agronomically important genes would be useful to improve Korean soybean improvement.

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Examination of Parameters Affecting Polymerase Chain Reaction in Studying RAPD (PCR에 의한 RAPD marker들의 증폭에 영향을 주는 조건들에 대한 고찰)

  • Yoon, Cheol-Sik
    • The Korean Journal of Mycology
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    • v.20 no.4
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    • pp.315-323
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    • 1992
  • The effects of several parameters on PCR amplification in using RAPD were studied. The results of this study suggest that approximately 15 ng of genomic DNA in $20\;{\mu}l$ of reaction mixture results in discrete and reproducible PCR products. In addition, the results indicate that concentration or amounts of reaction components studied are highly inter-dependent in their effects, and RNA can interfere severely with PCR amplification. Suitable concentrations or amounts of reaction components were found to be 30 ng of 10-mer primer, $200\;{\mu}M$ of dNTP, 0.001% gelatin 1.5 mM $MgCl_2$, 10 mM Tris-Cl (pH 8.8), 50 mM KCl, 0.1% Triton X-100, 2 units of Taq DNA polymerase, and 15 ng of RNase-treated genomic DNA in $25\;{\mu}l$ of reaction mixture.

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The Investigation of Gene Flows in Artificial Pollination between GM Rice and its Wild Relatives by RAPD Analysis (RAPD PCR에 의한 GM벼의 야생 근연종 벼로의 유전자 전이 분석법)

  • Kim, Yoon-Sik;Kim, Hyun-Soon;Joung, Hyouk;Jeon, Jae-Heung
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.612-616
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    • 2006
  • In recent years, there has been increasing concerns in gene flow from GM crops to wild or weedy relatives as a potential risk in the commercialization of GM crops. To access the possibility of the environmental impacts by GM rice, small-scale experiments of gene transfer were carried out. Herbicide and drought stress resistant GM rice and non-GM rice Nakdongbyeo, wild rice Oryza nivara, and weedy rice Sharebyeo were used for artificial pollination experiments and bar gene was used as a tractable marker after pollination. The harvested putative hybrid seeds after artificial pollination were germinated and true hybrid plants were selected by basta treatment. The hybrid plants were verified again by PCR amplification of bar and trehalose-6-phosphate phosphatase (TPP) genes and RAPD PCR analysis.

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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Identification of Hanwoo Meat by DNA Analysis (DNA 분석법에 의한 한우고기 판별)

  • Oh, Hong-Rock;Lee, Chang-Soo;Sang, Byung-Chan;Song, Kwang-Taek
    • Korean Journal of Agricultural Science
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    • v.33 no.1
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    • pp.1-10
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    • 2006
  • This study was deal with the development of breed-specific DNA marker which is able to identify Hanwoo and European cattle breeds(Non-Hanwoo) meat. Genetic differentiation between Korean cattle(Hanwoo) and European cattle breeds was examined by Random Amplified Polymorphic DNA(RAPD) analysis. The RAPD patterns were identical among Non-Hanwoo, such as Holstein, Hereford, Aberdeen Angus, Brown Swiss, Limousin or Simmental, but the above pattern was different from that of Hanwoo. All bands detected in the Hanwoo samples were observed in Non-Hanwoo cattle samples, but one of the common bands found in samples was not detected in the Hanwoo samples. The band(1.4kb) may be useful as a marker for identifying a meat of Hanwoo from imported cattle meat. Actually, the detection of the DNA marker was tested by DNA analysis with 929 samples which were prepared from bloods of 673 Hanwoo cattles and 141 Holstein cattles, from 115 imported cattle meats. The DNA marker was absent in 644 of 673 Hanwoo cattles (96%) but present in 245 of 256 Non-Hanwoo cattles (95%). These results show that the DNA marker is effective to characterize Hanwoo and Non-Hanwoo meat by its detection. This DNA marker, however, was not useful in detecting unwanted crossbreeding between two cattle breeds, because the band pattern in hybrid cattle shows one of two band patterns in Hanwoo and Non-Hanwoo.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Kacip Fatimah (Labisia pumila Benth & Hook f) collected from Melaka and Negeri Sembilan States of Malaysia

  • Bhore, Subhash J.;Nurul, A.H.;Shah, Farida H.
    • Journal of Forest and Environmental Science
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    • v.25 no.2
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    • pp.93-100
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    • 2009
  • In Malaysia, Labisia pumila Benth & Hook f, popularly known as 'Kacip Fatimah' has been used traditionally to treat various elements of the woman's health in Malay community. The objective of this study was to develop randomly amplified polymorphic DNA (RAPD) based DNA markers for the identification of L. pumila and to distinguish its three varieties from each other. Total DNA from nine accessions of L. pumila was extracted by CTAB method and polymerase chain reactions (PCR) were carried out to amplify the segments of DNA using different primers to develop DNA barcode using RAPD technique. To find out variety-specific DNA marker/s, twenty different 10-mer primer sequences with annealing temperature from 36-$40^{\circ}C$ were evaluated in triplicate. Out of 20 random primers, two primers (OPA-1 and OPA-2/A10) were selected which produced reliable RAPD band patterns. To have DNA based handle, two RAPD amplification products were cloned and sequenced to determine the identity of the DNA. RAPD analysis using two random primers generated 72 discrete bands ranging in size 200 bp-3,000 bp. Fifty nine of these were polymorphic loci (82%) and thirteen were non-polymorphic loci (18%). A total of 32 bands polymorphic loci (72%) were amplified with primer OPA-1 and analyzed by cluster analysis and UPGMA (Unweighted Pair Group Method with Arithmetic) to present a dendogram depicting the degree of genetic relationship among nine accessions of L. pumila. Our results shows the reasonable genetic diversity among the L. pumila varieties and within varieties; and two RAPD marker sequences obtained could be used to identify L. pumila at species level.

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