• Title/Summary/Keyword: RAPD primer analysis

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The Specific Probes Confirming the Genomic DNA of Tricholoma matsutake in Korea (송이의 Genomic DNA에 특이적인 Probe)

  • Lee, Sang-Sun;Hong, Sung-Woon;Chung, Hung-Chae;Sung, Chang-Kun;Kim, Jae-Hun;Ka, Kang-Hyeon;Kim, Hyun-Joong
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.20-26
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    • 1999
  • The specific DNA band appeared in PCR-RAPD analysis using OPO-2 primer was a very important for the researching Korean pine-mushrooms, Tricholoma matsutake. This DNA band, sequenced to be the 770 base pairs, existed as only a single copy in the whole genomic DNA's of Korean pine-mushrooms. However, this band was not presenting from the PCR-RAPD bands of other ectomycorrhyzal fungi reacted with the OPO-2 primer or the dot blots. Also, this DNA sequence was not matched with those of the other genes known by NCBI and had low homology together with sequence of other proteins compared. Those results suggested that the specific DNA band can be used as probe for identification of T. matsutake and might be related to the informations rather than the gene for the proteins with analysis of protein sequence translated from the DNA sequence.

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Genetic Diversity of Glutinous Rice Collections Based on Agronomic Traits and RAPDs (작물학적 형질과 RAPD에 의한 찰벼 수집종의 유전적 다양성)

  • Kim Guk-Hwan;Kim Hong-Sig;Lee Seok-Young;Chung Bong-Hwan;Song Beom-Heon;Cho Yong-Gu
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.3
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    • pp.212-220
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    • 2005
  • One hundred eleven glutinous rice collections from seven countries were evaluated for genetic diversity based on agronomic traits and RAPD analysis. Twelve agronomic traits including yield components amy-lose content, alkali digest value were used to clarify the genetic relationships among glutinous rice collections. Glutinous rice collections were classified into 4 groups and early maturings Korean landraces and high amylose indica belonged to group I with RAPD analysis, 15 primers selected for polymorphic bands generated 117 bands and 81 bands $(69.2\%)$ showed polymorphism. The number of amplified bands per primer ranged from 5 to 11, with the average number of bands of 7.8. With the similarity value of 0.78 in dendrogram derived from the cluster analysis based on RAPDs, glutinous rice collections were classified into 9 groups. Seventy-seven percent of the collections were classified into group I that is the largest one, while the others $(23\%)$ were distributed to group Il-IX. Group I included most indica type rices and early ripening collections, while the small groups of III-IX included most of the Korean collections.

RAPD Analysis of DNA Polymorphism and Genetic Species-Specificity Using PCR Technique in the Marine Microalgae

  • Yoon, Jong-Man;Chang, Kye-Nam
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.487-488
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    • 2000
  • Genomic DNA was isolated from the marine microalgae representing genetic characteristics and genomic polymorphisms by polymerase chain reaction amplification of DNA as arbitrary primers. The electrophoretc analysis of PCR-RAPD products showed hig levels of variation between different genus and little variation between different species. Outer of these primers, 6 generated 248 highly reproducible RAPD markers, producing almost seven polymorphic bands per primers. The degree of similarity frequency between Chaetoceros gracilis and Chaetoceros calcitrans species showed 90% as calculated by sharing analysis. The RAPD polymorphism generated by this primer may be used as a genetic marker for genus or species identification in important marine microalgae. (omitted)

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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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Intrapecific Relationship of Rehmannia glutinosa Lines Collected from Korea, Japan and China by RAPD Analysis (RAPD 방법을 이용한 국내외 수집 지황(地黃)의 유연 관계 분석)

  • Kim, Jong-Yeob;Choi, Sun-Young;Choo, Beng-Gil;Ryu, Jeom-Ho;Kwon, Tae-Ho;Oh, Dong-Hun
    • Korean Journal of Medicinal Crop Science
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    • v.8 no.3
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    • pp.266-273
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    • 2000
  • The optimal conditions of PCR components for the random amplification of genomic DNA were $20\;ng/20{\mu}l$ in template DNAs, 250 mM in dNTP, 10 pM in primer $1.0unit/20{\mu}l$ in Taq DNA polymerase respectively with the annealing temperature at $36^{\circ}C$, respectively. Twelve local lines were divided into 3 groups by the coefficients of 107 polymophic bands by Jaccard and Nei. The coefficients value of group I including Chongup # 1, Seochon # 1, Andong # 1, Chinan # 1, and Danyang # 1 ranged from 0.27 to 0.05 and those of group II including Suwon # 2, Chunchon # 1, Japan # 3, Danyang#2 and $F_1$ (Variety Jihwang $1{\times}$ Seohchon) ranged from 0.29 to 0.11. While, Jihwang 1 originated from China and Japan # 1 in group III showed a distant genetic relationship to Korean local lines.

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Identification of Genetic Markers for Korean Native Cattle (Hanwoo) by RAPD Analysis

  • Yeo Jung Sou;Lee Ji Sun;Lee Chang Hee;Jung Young Ja;Nam Doo Hyun
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.1
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    • pp.23-26
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    • 2000
  • In order to develop the specific genetic marker for Korean native cattle (Hanwoo), randomly amplified polymorphic DNA (RAPD) analysis of 6 different cattle breeds was attempted by using 38 decamer primers. In comparison of RAPD patterns, two distinctive DNA bands specific for Hanwoo were detected. One was 296 bp of DNA fragment found to be specific only for female Hanwoo when primer GTCCACACGG was employed. In individual analysis of this RAPD marker was observed only in female individuals with the possibility of $85.3\%$. The other was 521 bp of RAPD marker amplified using TCGGCGATAG and AGCCAGCGAA primers, which showed $83.0\%$ of genetic frequency in 85 male and 68 female individuals tested. Nucleotide sequencing of these genetic markers revealed that 296 bp marker has a short micro satellite-like sequence, ACCACCACAC, and a tandem repeat sequence of microsatellite GAAAAATG in the determined sequence. Two distinctive tandem repeats of microsatellite sequences, MC and GAAGA, were also appeared in 521 bp DNA marker. In BLAST search, any gene having high homology with these markers was not found.

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Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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Relationship with botanical characteristics and RAPD analysis of maize inbred lines with tillers (분얼형 옥수수의 식물학적 특성과 RAPD 분석에 의한 근연관계)

  • Kim, Chol-Min;Lee, Hee-Bong;Choi, Hyun-Gu;Jung, Jae-Young;Kim, Gi-Hoon
    • Korean Journal of Agricultural Science
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    • v.28 no.1
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    • pp.8-17
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    • 2001
  • This study was carried out to obtain a basic intonation for the development of a new com hybird with tillers. Materials used in this study were 20 lines having three to four tillers per plant including the PI213749 U.S. line with non-tillers. These 20 lines were compared for the botanical characteristics and genetic distances were measured using RAPD analysis. Flowering date of the K15 was very earlier, while the K07 was very late in flowering date. Stem height and ear height were similar except for K04 and the K15. K06, K13 and K19 lines were appeared to be resistant to lodging due to decreased ear height. Number of tillers per plant of lines used were shown three to five on average. K09 showed the highest kernel yield, while the K08 was low. Among characteristics measured tiller per plant and flowering dates, and silking dates showed a positive correlation, while 100 kernels weight, flowering date and ear height were shown a negative with tillers per plant. A total of 17 bands by RAPD analysis using four per primer were appeared and these lines were classified into three groups, especially the third group could be classified into of four sub-groups.

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Development of a psychrophilic-SCAR marker for Pleurotus eryngii (큰느타리버섯의 저온적응성 형질에 관련된 SCAR Marker 개발)

  • Kim, Su Chul;Hwang, Hye Sung;Cho, Yun Jun;Kim, Hye Su;Ryu, Jae-San;Cho, Soo Jeong
    • Journal of Mushroom
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    • v.11 no.3
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    • pp.171-176
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    • 2013
  • Genomic DNAs of psychrophilic strains of Pleurotus eryngii were analyzed by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S3 primers to develop the strain-specific DNA marker. A unique DNA fragment with the size of 480 bp was yielded by OP-S3 primer from the psychrophilic strain. A sequence characterized amplified region (SCAR) marker, designated as OP-S3-1, was designed on the basis of the determined sequence. The PCR analysis with the OP-S3-1 primer showed that this SCAR marker can clearly distinguish the psychrophilic strains from the control strains.

Genotyping Based on Polymerase Chain Reaction of Enterobacter sakazakii Isolates from Powdered Infant Foods

  • Choi, Suk-Ho;Choi, Jae-Won;Lee, Seung-Bae
    • Food Science and Biotechnology
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    • v.17 no.6
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    • pp.1171-1177
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    • 2008
  • This study was undertaken to classify Enterobacter sakazakii isolates from 13 powdered infant formula products, 25 powdered weaning diet products, and 33 weaning diet ingredients on polymerse chain reaction (PCR) methods. The numbers of the isolates from 1 powdered infant formula product, 7 powdered weaning diet products, and 6 weaning diet ingredients were 1, 14, and 8, respectively. The contaminated ingredients were 1 rice powder, 2 millet powders, 2 vegetable powders, and 1 fruit and vegetable premix. PCR with the primer of repetitive extragenic palindromic element (REP-PCR) and random amplification of polymorphic DNA(RAPD) were effective in discriminating among the isolates, but tRNA-PCR and PCR with the primer of l6S-23S internal transcribed spacer (ITS-PCR) were not. Some of E. sakazakii isolates from vegetable powders, fruit and vegetable premix, and millets powders were classified into the clonal groups based on the DNA patterns in the REP-PCR and RAPD analysis. A close genetic relationship among the isolates from some of the powdered weaning diet products and the rice powder was also detected in the cluster analysis based on the DNA patterns in RAPD.