• Title/Summary/Keyword: random primer

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Identification of RAPD markers linked to sex determination in guggal [Commiphora wightii (Arnott.)] Bhandari

  • Samantaray, Sanghamitra;Geetha, K.A.;Hidayath, K.P.;Maiti, Satyabrata
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.95-99
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    • 2010
  • Decamer RAPD primers were tested on dioeceious and hermaphrodite plants of Commiphora wightii to identify sex-specific molecular markers. Sixty different random decamer primers were screened out of which only three primers were found to be associated with sex expression. A ~1,280-bp fragment from the primer OPN06 was found to be present in all the female individuals. Another primer OPN 16 produced a unique ~400-bp amplification product in only hermaphrodite individuals. The third marker, OPA20 amplified a ~1,140-bp fragment from female and hermaphrodite DNAs, but failed to do so from the male plant DNAs.

Genetic Variability of Pleurotus ostreatus Monospore Isolates by Random Amplified Polymorphic DNA Analysis (DNA 다형성 분석에 의한 느타리버섯 단포자 분리주의 유전적 변이성)

  • Song, Yeong-Jae;Jeong, Mi-Jeong;Kim, Beom-Gi;Rho, Yeong-Deok;Ryu, Jin-Chang;Yoo, Young-Bok
    • The Korean Journal of Mycology
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    • v.24 no.3 s.78
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    • pp.186-205
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    • 1996
  • This study was carried out to obtain data concerning the genetic variability of Pleurotus ostreatus. Monospores of P. ostreatus were isolated and cultured to estimate differences in the rate of mycelial growth and genetic similarity among the isolates. Although the overall growth rates were slow compared to their parental dikaryon except 2-MI 4, significant differences in the rate of mycelial growth were observed among isolates. Random amplified polymorphic DNA (RAPD) analysis using twenty six random primers showed 345 polymorphic DNA bands from 35 monospore isolates and their parental dikaryon. DNA bands ranged from 0.1 to 4.0 Kb in size. Most various polymorphic DNA bands within monospore isolates were obtained when we used J (OPA-01) or W (OPB-04). The 36-MI 103 showed totally different band patterns from those of the others. RAPD analysis revealed that there is approximatly 75% genetic similarity between monospore isolates and their parental dikaryon except 36-MI 103, which showed only 49% genetic similarity. In addition, genetic similarity degrees were classified into four groups: I (parental dikaryon), II (fast growing type), III (moderate growing type), and IV (slow growing type). However, there is no correlation between mating type and mycelial growth rates.

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Genetic variation of local varieties and mutants groups induced by gamma ray in Hypsizigus marmoreus (느티만가닥버섯의 재래종과 감마선 돌연변이체들의 유전적 변이)

  • Kim, Jong-Bong;Yu, Dong-Won
    • Journal of Mushroom
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    • v.12 no.3
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    • pp.181-186
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    • 2014
  • This research was carried out to analyze the genetic variation of 18 wild strain, 2 breed varieties and 20 mutants of Hypsizygus marmoreus by random amplification of polymorphic DNA(RAPD). Also, 3 strains of Lyophyllum decartes and 1 strain of Lyophyllum shimeji were used. These mushrooms were collected from korea, china, Taiwan and Japan. Spores of H. marmoreus JV-2 strain were irradiated by gamma ray for mutagenesis. 40 kind of primers were used for this reaserch. Number of reaction primer were 31. Electrophorectic patterns of RAPD showed genetic variation. In phylogenetic tree, they were divided into seven group. Discriminative differences were observed between wild strain and mutants in H. marmoreus. These results might suggest that these primers and gamma ray irradiation of spores were useful tools for developing new strain for mushroom.

Genetic Variation and Identification of RAPD Markers from Some Garlic Cultivars in Korea and Mongolia (한국과 몽고 일부 재배마늘의 유전적 변이와 재배종 특이적 RAPD 마커의 탐색)

  • Bae, Seong-Kuk;Jung, Eun-A;Kwon, Soon-Tae
    • Korean Journal of Plant Resources
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    • v.23 no.5
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    • pp.458-464
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    • 2010
  • Twelve garlic cultivars collected from Korea and Mongolia were evaluated genetic similarity and diversity by RAPD method using oligo-nucleotide random primers. Genomic DNA isolated from twelve garlic cultivars were amplified by polymerase chain reaction using 143 primers, and 55 primers showed polymorphic DNA bands. Among a total of 187 bands amplified by 55 primers, 128 polymorphic bands were subjected to analysis for genetic relationship of garlic cultivars. Garlic cultivars were classified into three groups, such as group-I corresponded to Euiseong, Seosan, Samchuk and Yecheon-A, Yechun-B, Euiseong-norang, Jeongsun, Namdo, Yookback and Danyang cultivars, and group-II to Mongolia and group-III to Daeseo cultivars. Thirty DNA bands showing unique specificity to the specific cultivars are likely to be useful for identification of garlic local cultivars as DNA markers.

Genetic Identity between Bhadawari and Murrah Breeds of Indian Buffaloes (Bubalus bubalis) Using RAPD-PCR

  • Saifi, H.W.;Bhushan, Bharat;Kumar, Sanjeev;Kumar, Pushpendra;Patra, B.N.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.603-607
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    • 2004
  • Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out with a battery of 11 random decamer primers to study band frequency (BF), genetic identity index (I) and mean average percentage difference (MAPD) between Bhadawari and Murrah breeds of buffalo. The primers OPA04 and BG15 resolved a band of 460 bp, which was present only in animals of Bhadawari breed. Whereas, the primers OPA14, BG27 and BG28 produced Murrah specific fragments of sizes 730 bp and 1,230 bp, respectively. The estimate of genetic identity index was highest (0.845) with the primer OPA01 and the lowest (0.479) with the primer BG27. The genetic identity index pooled over the primers was 0.596${\pm}$0.037 between these two breeds. The highest MAPD estimate (53.9) between the two breeds was obtained with the primer BG27 and the lowest (14.3) with the primer OPA01. It might be concluded that the genetic identity index between these two breeds calculated on the basis of BF showed moderate level of genetic identity with the primers employed. MAPD calculated on the basis of uncommon bands also demonstrated lower to medium level of genetic difference between Bhadawari and Murrah breeds of buffalo.

Limits of Direct PCR Amplification from Seaweeds Using Arbitrary and ITS Primers (해조류로부터 Arbitrary 및 ITS Primer들을 사용한 직접 PCR 유전자 증폭반응의 한계)

  • 김용국;진형주;박선미;진덕희;홍용기
    • Journal of Life Science
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    • v.9 no.1
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    • pp.15-21
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    • 1999
  • The random amplified ploymorphic DNAs (RAPD) assay is a simple and useful tool in identification of appropriate genetic markers, that requires no knowledge of target DNA sequence. RAPD products were generated directly from seaweed tissues, without prior nucleic acid extraction, of Porphyra yezoensis, Ulva pertusa and Undaria pinnatifida. The nuclear rDNA internal transcribed spacer (ITS) fragment however was not amplified directly from the seaweed tissues. Using DNA extracted by the LiCl method, both the ITS and RAPD's have been amplified by the polymerase chain reaction. RAPD of P yezoensis, thallus (n) and conchocelis (2n) produced lots of different polymorphic bands (36-50$\%$) depending on the arbitrary primers used. Difference was also observed between direct tissues amplification and DNA extracts amplification (53-57$\%$). Thus it is important to use the same ploidy of tissue for DNA extraction and as a RAPD template.

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Characteristics Comparisons of Edwardsiella tarda Isolated from Cultured Olive Flounder and Eel (양식넙치와 뱀장어에서 분리된 Edwardsiella tarda의 특성 비교)

  • Kim, Eunheui
    • Journal of fish pathology
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    • v.34 no.1
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    • pp.31-38
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    • 2021
  • The objective of this study was to determine comparative biochemical characteristics and RAPD (random amplified polymorphic DNA) profiles of 18 strains of Edwardsiella tarda isolated from cultured olive flounder (Paralichthys olivaceus) and eel (Anguilla spp) that showed diseases between 1996 and 2010 in Korea. In terms of biochemical properties, they showed four patterns with differences in citrate degradation and production of H2S and indole. All strains were identified as E. tarda. Characteristics of isolates were not classified according to their host. As a result of PCR with E. tarda-specific primer, EDtT, the same band of about 270 bp was detected in all 18 isolates. However, no specific band was detected in type strains of E. tarda or Edwardsiella ictaluri. As a result of RAPD PCR performed with primer No. 5 and No. 6 of a Ready-To-Go-RAPD kit, the band profile showed clear differences among isolates of olive flounder, isolates of eel, and E. tarda and E. ictaluri type strains. It was possible to organize their characteristics according to the origin of host with possibility to develop tools for pathogen monitoring.

Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis (REP-PCR을 이용한 국내 사람과 동물유래 Staphylococcus aureus 분리주의 Molecular Typing)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.60-66
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    • 2005
  • To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.

In vitro propagation of oil palm (Elaeis guineensis Jacq.) clones through somatic embryogenesis and analysis of somaclonal variation by RAPD (체세포배발생을 통한 오일팜나무(Elaeis guineensis Jacq.) 클론의 기내증식 및 RAPD를 이용한 체세포변이의 검정)

  • Ahn, In-Suk;Park, Hye-Rim;Son, Sung-Ho
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.196-204
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    • 2012
  • This study was carried out to develop reliable systems for somatic embryogenesis in oil palm tree (Elaeis guineensis Jacq.), and to verify the somaclonal variants by RAPD analysis. Embryogenic callus was induced successfully on modified half-strength MS medium containing $NaH_2PO_4{\cdot}2H_2O$ and casein. Embryogenic callus was further developed to somatic embryo mass (SEM), which is very hard and bonded tightly each other. Plantlets were proliferated when SEM was cultured on modified MS medium containing half strength $NH_4NO_3$, casein and L-ascorbic acid. Plantlets were transplanted into pots containing artificial soils. When RAPD analysis was conducted using randomly selected 95 in vitro plantlets and 19 random primers, somaclonal variation was detected using BNR35 primer. There was missing band around 1 kb in #22, #28, #35, and #77 plantlets. In addition, bands obtained from #28, #35, and #77 was much stronger than other normal bands. The blast results at NCBI revealed that somaclonal variation observed in this study was related to chloroplast genome of oil palm. The results also revealed that oil palm reproduction system through somatic embryogenesis is quite reliable and early detection of somaclonal variants seem to be possible at in vitro stage by RAPD analysis.

Differences by RAPD-PCR Analysis within and between Rockfish (Sebastes schlegeli) Populations from the Yellow Sea and the Southern Sea in Korea (황해 및 남해산 조피볼락 (Sebastes schlegeli) 개체군 사이의 RAPD-PCR 분석에 의한 차이)

  • Yoon, Jong-Man;Kim, Jong-feon
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.359-369
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    • 2001
  • Polymerase chain reaction (PCR) amplification of DNA as 30 different arbitrary primers and random amplified polymorphic DNAs (RAPD) analysis were performed on genomic DNA extracted from the blood of the marine rockfish (Sebastes schlegeli) from the Yellow Sea and the Southern Sea. The unique properties of the genomic DNA were used to investigate the features of the population dynamics and origins of the species. Out of 30 primers, seven generated 207 highly reproducible RAPD polymorphic products, producing approximately 2.7 polymorphic bands per primer. About 67.4% of total amplified products (307) were either polymorphic (207) to rockfish. The degree of similarity varied from 0.22 to 0.63 as calculated by bandsharing analysis. Also, the average level of bandsharing was 0.39$\pm$0.02 within the rockfish strains. The electrophoretic analysis of RAPD-PCR products showed the relatively high levels if variation between different individuals in rockfish from the Yellow Sea. However, the RAPD outlines obtained with DNA of different rockfish strains from the Yellow Sea and the Southern Sea in Korea were very similar. Also, a small number of polymorphic bands were identified. Even if further analyses or more rockfish populations are required, this result implies RAPD analysis reflects genetic differences between the geographical strains of the rockfish.

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