• Title/Summary/Keyword: random amplified polymorphic DNA (RAPD) marker

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Random Amplified Polymorphic DNA-PCR Analysis for Identification of Bacillus anthracis (탄저균의 Random Amplified Polymorphic DNA-PCR 분석)

  • 김성주;박경현;김형태;조기승;김기천;최영길;박승환;이남택;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.56-60
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    • 2001
  • Molecular typing of Bacillus anthracis has been extremely difficult due to the lack of polymorphic DNA markers. Aiming to develop a DNA marker specific for Bacillus anthracis and to be able to discriminate this species from Bacillus genus, we applied the random amplified polymorphic DNA (RAPD)-PCR. We have identified B. anthracis from various Bacillus species. The analysis performed by RAPD clearly demonstrated substantial genetic variations among Bacillus species. This type of analysis is an easy, quick and highly discriminatory technique that may help in diagnosis of anthrax.

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Differentiation of Fusarium oxysporum f. sp. fragariae Isolates by Random amplified Polymorphic DNA (RAPD) Analysis. (Random Amplified Polymorphic DNA(RAPD)를 이용한 딸기 시들음병균(Fusarium oxysporum f. sp. fragariae)의 분류)

  • 현재욱;박원목
    • Korean Journal Plant Pathology
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    • v.12 no.1
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    • pp.41-46
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    • 1996
  • 본 실험은 이병 딸기의 조직에서 분리 동정된 시들음병균(Fusarium oxysporum f. sp. fragariae) 균주들의 유?거 변이를 random amplified polymorphic DNA(RAPD) marker들을 이용하여 조사하였다. 총 24개의 딸기 시들음병 균주들의 DNA를 주형으로 하여 16개의 random 10-mer primer들을 사용하여 증폭시킨 결과 총 231개의 marker들을 이용하여 유전적 변이를 조사해 본 결과 크게 RAPD I과 RAPD II의 2개 그룹으로 나눌 수 있었다. RAPD I그룹에 속하는 균주는 VCG A에 속하는 Y1, K1, K2, K3, K4, N2, N3, N4-1, N6-1, N6-2, N8, N9, N10, M1-2-1 균주, VCG B에 속하는 M4-1 균주 그리고 VCG C에 속하는 N1, Y2 균주들이었고, RAPD II그룹에는 VCG B에 속하는 M1-1, M2-2-1, M2-4-2, M3-2, M3-3-2 균주와 VCG D에 속하는 N1 1 균주가 속하였다. 이들 2그룹 간에는 31%의 유사성이 있었다.

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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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Discrimination of the Genus Leontopodium Species (Gentianales: Asteraceae) Based on RAPD

  • Jeon, Mi Gyeong;Choi, Kang Jun;Kim, Ji Young
    • Journal of Forest and Environmental Science
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    • v.31 no.1
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    • pp.68-71
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    • 2015
  • Korean L. leiolepis of the genus Leontopodium could be discriminate from the foreign L. alpinum using random amplified polymorphic DNA (RAPD). Among the 12 URP markers used for the detection, the URP-5 marker and the URP-7 marker detected polymorphic DNA bands, ranging from 400-1000 bp in the size of amplified DNA fragments.

Genomic Polymorphisms of Genome DNA by Polymerase Chain Reaction-RAPD Analysis Using Arbitrary Primers in Rainbow Trout (PCR-RAPD 기법에 의한 무지개송어 Genome DNA 의 다형현상)

  • Yoon, J.M.
    • Korean Journal of Animal Reproduction
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    • v.23 no.4
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    • pp.303-311
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    • 1999
  • Nuclear DNA was isolated from the sperm cells representing genetic characteristics and genomic polymorphisms of rainbow trout by polymerase chain reaction(PCR) amplification of DNA using arbitrary primers. Genomic DNA fingerprints were generated from rainbow trout sperm DNA by polymerase chain reaction amplification using 20 arbitrary decamers as primers. Out of these primers, 4 generated 17 highly reproducible RAPD markers, producing almost six polymorphic bands per primers. Four of 6 primers tested generated amplified fragments which were polymorphic between different individuals. Polymorphic DNA fragments were reproducibly amplified from independent DNA preparations made from individuals. Rainbow trout was distinctly observed 3 specific DNA markers (2. 3, 2.0 and 1.3kb) in bandsharing. Individual fragments generated using the same arbitrary primer, demonstrated that a single primer detected at least three independent genomic polymorphisms in rainbow trout sperm DNA. The RAPD polymorphism generated by this primer may be used as a genetic marker for individual identification The RAPD-PCR technique has been shown to reveal informative polymorphism in many species of fish. The present results demonstrate that RAPD markers are abundant, reproducible and provide a basis for future gene mapping and MAS in these important aquaculture species using RAPD polymorphic markers. It is concluded that RAPD polymorphisms are useful as genetic markers for fish breed differentiation.

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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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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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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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Genetic Variation Based on Random Amplified Polymorphic DNA (RAPD) and Internal Transcribed Spacer (ITS) Region Sequences in Lepista nuda (RAPD와 ITS 영역에 의한 민자주방망이 버섯의 유전적 변이)

  • Lee, Yang Suk;Kim, Nam Woo;Kim, Jong Bong
    • Journal of Life Science
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    • v.22 no.11
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    • pp.1470-1476
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    • 2012
  • A genetic variation of Lepista nuda and two genus Lepista species (L. irina and L. sordida) were analyzed by random amplified polymorphic DNA (RAPD) and internal transcribed spacer (ITS) sequence analysis. In the resulting RAPD analysis, 22 out of 40 random primers amplified polymorphic RAPD fragment patterns, the amplified bands were 355, and DNA fragment sizes were 200-400bp. Intraspecific genetic dissimilarity of the 10 L. nuda strains were calculated to range from 0% to 21.60%, L. sordida from 16.93% to 24.82%, L. irina were 20.62% to 25.54%, and intraspecific genetic dissimilarity of L. sordida and L. irina was 23.49%. The 673 base pairs were sequenced during the analysis of the ITS I and II region; six L. nuda strains intraspecific genetic dissimilarities ranged from 1.58% to 11.47%, L. nuda and L. sordida from 3.83% to 12.88%, L. nuda and L. irina from 7.11% to 15.61%, and intra-specific genetic variation between L. sordida and L. irina was 4.79%. The findings showed that RAPD and ITS sequencing could be used for developing molecular genetic markers and screening of unidentified genus Lepista species.

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.