Investigations of Genetic Variation Between Olive (Olea europaea L.) Cultivars Using Arbitrarily Primed Polymerase Chain Reaction (AP-PCR)

2003 ◽  
Vol 58 (11-12) ◽  
pp. 837-842 ◽  
Author(s):  
Feray Kockar ◽  
Rahsan Ilıkcı
Keyword(s):  
Genetic Relationship ◽  
Genomic Dna ◽  
Pcr Amplification ◽  
Dna Fingerprint ◽  
Factors Affecting ◽  
Genomic Dna Isolation ◽  
Amplification Protocol ◽  
Olive Cultivars ◽  
Arbitrarily Primed Pcr ◽  
Relationship Of

Abstract Characterization and selection of olive clones for the production of olive oil is essential in Turkey because of its profitable exploitation. AP-PCR (Arbitrarily-Primed PCR) is a technique that can distinguish the genetic relationship among plant species and other organisms. In this study, AP-PCR approach was used in order to determine the genetic relationship of different six olive clones. The purity of DNA is one of the most important factors affecting the product of the AP-PCR method. In this respect, modified genomic DNA isolation procedure from Oleae europaea clones was developed so that this procedure can be used to obtain plant genomic DNA from diverse aromatic plants, which produce essential oils and secondary metabolites. By following the optimized AP-PCR amplification protocol, unique DNA fingerprint profiles for each olive clone were produced. AP-PCR-generated unique DNA fingerprint profiles can be used in the identification, distribution and diversity of various olive cultivars.

scholarly journals Optimizing the pure genomic DNA isolation procedure for Plectranthus amboinicus DNA – A prerequisite for further genomic Studies

2017 ◽  
Vol 2 (4) ◽  
Author(s):  
S. Manikandan ◽  
S. Ansarali ◽  
G.M. Alagu Lakshmanan
Keyword(s):  
Dna Barcoding ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Leaf Tissue ◽  
Isolation Procedure ◽  
Modified Method ◽  
Genomic Dna Isolation ◽  
Young Leaves ◽  
Plectranthus Amboinicus

The DNA isolation procedure for different plant groups have been studied and standardized.  The isolation of pure genomic DNA is the most essential component for any type of molecular studies.  The present work is aimed to identify suitable DNA markers for the amplification of P.amboinicus  DNA  becomes a great hurdle  for DNA barcoding studies carried out by rbcL and matK primers Used in the members of Lamiaceae. To solve this problem, The DNA was extracted by three methods from fresh young leaf tissue of P.amboinicus. After the evaluationthe outcome of these methods, one most suitable modified method was selected for isolating DNA from young leaves of P.amboinicus and selected for suitable DNA barcoding markers for PCR amplification. The quality and quantity of DNAs are a prerequisite for genetic studies for a variety of plants including P.amboinicus. The quantity and quality of the DNA extracted by this method wasused for suitable DNA barcoding markers selection.

scholarly journals Optimizing Genomic DNA Isolation and PCR Amplification For Pasak Bumi (Eurycoma longifolia)

2019 ◽  
Vol 1 (2) ◽  
pp. 30
Author(s):  
Arida Susilowati ◽  
Henti Hendalastuti Rachmat ◽  
Ahmad Baiquni Rangkuti ◽  
Deni Elfiati ◽  
Ami Ambarwati
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Pcr Amplification ◽  
Eurycoma Longifolia ◽  
Genomic Dna Isolation

.

scholarly journals mecA-Gene Detection and MDR Profile of S. Aureus Isolates From Patients Attending The Referral Hospitals of Amhara Reginal State, Ethiopia

2021 ◽  
Author(s):  
Feleke Moges ◽  
Tadele Tamiru ◽  
Azanaw Amare ◽  
Getachew Mengistu ◽  
Wondwossen Abebe ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Meca Gene ◽  
Surrogate Marker ◽  
Pcr Amplification ◽  
Resistance Rate ◽  
Molecular Techniques ◽  
High Rate ◽  
Amhara Region ◽  
Genomic Dna Isolation ◽  
Referral Hospitals

Abstract Background: Staphylococcus aureus causes different types of human infections, and has an ability to develop resistance to many antibiotics. There is scarcity of data on mecA gene and MDR profiles of this organism in developing countries, like Ethiopia. The aim of the present study is therefore, to investigate MDR profiles and mecA-gene profile of S. aureus from Referral Hospitals of Amhara Reginal State. Methods: Of the total of 110 isolates collected from Amhara Region Referral Hospitals, 70 MDR isolates were further processed for isolation of S. aureus mecA gene. Genomic DNA was isolated using SIGMA ALDRICH genomic DNA isolation Kit for Gram positive bacteria. Amplification of S. aureus mecA gene was done using amplicon size of 533 bp. Agarose gel electrophoresis was prepared with 1.5% agarose by TAE solvent and 0.5μg/mL of ethidium bromide. Electrophoresis was carried out for 1 hour and a half (7 Volts/ cm²) in 1X tris acetate EDTA (TAE) buffer. Data were analysed by using descriptive statistics.Results: Majority of the isolates were recovered from patients age less than 5 years, 51 (36.7%) and least from age greater than 60 years, 6 (4.3%). Most of the isolates were from blood, 61 (43.9%), followed by wound, 45 (32.4%). High resistance rate was observed in penicillin 81 (73.6%), followed by cotrimoxazole 78 (70.9%), ceftriaxone 76 (69%), erythromycin 66 (60%) and tetracycline 65 (59.1%). Phenotypically, considering cefoxitin as a surrogate marker, 38 (34.5%) of the isolates were methicillin resistant. The overall MDR isolates were 92 (83.6%). The PCR amplification result of mecA gene was 14 (20%).Conclusions and recommendations: There are high rate of MDR and MRSA isolates of S. aureus. PCR amplification result indicates 20% of MRSA isolates are mecA gene producers. Largescale study for detection of MDR strains of S. aureus including MRSA using molecular techniques should be encouraged in Amhara region.

scholarly journals Comparative Analysis of the Genomic DNA Isolation Methods on Inula sp. (Asteraceae)

2016 ◽  
Vol 8 (4) ◽  
pp. 451-455
Author(s):  
Emre SEVİNDİK ◽  
Fatih COŞKUN ◽  
Zehra Tuğba MURATHAN ◽  
Mehmet Yavuz PAKSOY ◽  
Veysel UZUN
Keyword(s):  
Genomic Dna ◽  
Dna Isolation ◽  
Low Cost ◽  
Pcr Amplification ◽  
Isoamyl Alcohol ◽  
Dna Amount ◽  
Genomic Dna Isolation ◽  
Asteraceae Family ◽  
Positive Results ◽  
Commercial Kit

Simple, fast, low-cost and high throughput protocols are required for DNA isolation of plant species. In this study, phenol chloroform isoamyl alcohol and commercial (Sigma) DNA isolation kit methods were applied on some Inula species that belong to Asteraceae family. Genomic DNA amounts, A260, A280, A260/A230 and purity degrees (A260/A280) that were obtained through both methods were measured through electrophoresis and spectrophotometer. Additionally, PCR amplification was realized by primer pairs specific to nrDNA ITS, cpDNA ndhF (972F-1603R) and trnL-F regions. Results showed that maximum genomic DNA in nanograms obtained by phenol chloroform isoamyl alcohol method. The study also revealed that I. macrocephala had the maximum DNA and I. heterolepis had the minimum DNA amount. A260/A280 purity degrees showed that the highest and lowest purity in gDNAs obtained through phenol-choloform isoamyl alcohol method were in I.aucheriana and I. salicina, respectively. The highest and lowest purity degrees of gDNAs obtained through commercial kit was observed in I. fragilis and I. macrocephala samples, respectively. PCR amplification results showed that while band profiles of each three regions (ITS, trnL-F and ndhF) did not yield positive results in PCR amplifications using phenol-choloform isoamyl alcohol method; PCR band profiles obtained through commercial kit yielded positive results. As a result, it is fair to say that the relation of genomic DNA with PCR was found to be more efficient although the maximum amount of genomic DNA was obtained through phenol chloroform isoamyl alcohol method. 

scholarly journals Detection of Fusarium oxysporum f. sp. dianthi in Carnation Tissue by PCR Amplification of Transposon Insertions

1999 ◽  
Vol 89 (12) ◽  
pp. 1169-1175 ◽  
Author(s):  
Annalisa Chiocchetti ◽  
Ilaria Bernardo ◽  
Marie-Josée Daboussi ◽  
Angelo Garibaldi ◽  
M. Lodovica Gullino ◽  
...  
Keyword(s):  
Fusarium Oxysporum ◽  
Genomic Dna ◽  
Pcr Amplification ◽  
Dna Fingerprint ◽  
Genomic Dnas ◽  
Insertion Sites ◽  
Polymerase Chain ◽  
Transposon Insertions ◽  
Race 4

Strains of the carnation wilt pathogen, Fusarium oxysporum f. sp. dianthi, can be distinguished by DNA fingerprint patterns, using the fungal transposable elements Fot1 and impala as probes for Southern hybridization. The DNA fingerprints correspond to three groups of F. oxysporum f. sp. dianthi strains: the first group includes isolates of races 1 and 8; the second group includes isolates of races 2, 5 and 6; and the third group includes isolates of race 4. Genomic DNAs flanking race-associated insertion sites of Fot1 (from races 1, 2, and 8) or impala (from race 4) were amplified by the inverse polymerase chain reaction (PCR) technique. These regions were cloned and sequenced, and three sets of primers overlapping the 3′ or 5′ end of the transposon and its genomic insertion were designed. Using fungal genomic DNA as template in PCR experiments, primer pairs generated amplification products of 295, 564 and 1,315 bp, corresponding to races 1 and 8; races 2, 5, and 6; and race 4, respectively. When multiplex PCR was performed with genomic DNA belonging to races 1 and 8, 2, or 4, single amplimers were generated, allowing clear race determination of the isolate tested. PCR was successfully performed on DNA extracted from susceptible carnation cv. Indios infected with isolates representative of races 1, 2, 4, and 8.

scholarly journals KARAKTERISASI BEBERAPA STRAIN GURAMI Osphronemus gouramy Lac. MENGGUNAKAN MARKA RAPD

2014 ◽  
Vol 1 (1) ◽  
pp. 109
Author(s):  
Anisa Kartika Sari ◽  
Agus Nuryanto ◽  
Agus Hery Susanto
Keyword(s):  
Fish Species ◽  
Genetic Relationship ◽  
Genomic Dna ◽  
Rapd Markers ◽  
Dna Marker ◽  
Random Amplified Polymorphic Dna ◽  
Economic Value ◽  
Pcr Amplification ◽  
Sampling Technique ◽  
Survey Method

Giant gouramy, Osphronemus gouramy Lac. is a popular fish species in Indonesia, especially in Java and Sumatera as this freshwater fish species has a high economic value of stable price. Fish farmers in Bogor divide giant gouramy into six strains based on egg productivity, growth rate, and maximum weight of the adult. They are soang, jepang, blue saphire, paris, bastar, and porselin. These various strains lead to the need of study on the genetic relationship among them, which can be performed by the use of RAPD (Random Amplified Polymorphic DNA) marker. This study aims to determine primers producing consistent and polymorphic RAPD markers, determine specific RAPD markers, and know the genetic relationship among several giant gouramy strains. The strains used in this study are soang, jepang, and blue saphire. Survey method was applied employing purposive random sampling technique. Total genomic DNA was isolated using Genjettm genomic DNA purification kit (Fermentas), which was then used as template to amplify RAPD markers with primers OPA-07, OPA-09, OPA-11, OPA-20, OPAH-01, OPAH-08, OPAH-09, and OPAC-14. The variables examined were patterns and numbers of specific DNA fragments as the PCR amplification products. Selected primers were determined descriptively on the basis of specific DNA bands appearing on the agarose gel. Genetic diversity was predetermined by changing qualitative band pattern into quantitative binnary data. Genetic relationship was analyzed using cladistic method with PAUP software. The results showed that only five of the eight primers produce consistent and polymorphic RAPD markers, i.e. OPA-11, OPA-20, OPAH-1, OPAH-8, and OPAH-9. Specific RAPD markers which can be used to distinguish several gouramy strains are those amplified with OPA-20 of 786 bp, OPA-20 of 1,176 bp, OPAH-8 of 1,000 bp and OPAC-14 of 1,607 bp. Nervertheless, it was found that RAPD markers cannot be used to clearly determine genetic relationship among gouramy strains.

scholarly journals Assessment of the genetic relationship of Turkish olives (Olea europaea subsp. europaea) cultivars based on cpDNA trnL-F regions

2018 ◽  
Vol 77 (1) ◽  
pp. 88-92 ◽  
Author(s):  
Ergun Kaya ◽  
Recep Vatansever ◽  
Ertugrul Filiz
Keyword(s):  
Genetic Relationship ◽  
Olea Europaea ◽  
Phylogenetic Analyses ◽  
Olive Tree ◽  
Sequence Length ◽  
Mediterranean Countries ◽  
Olive Cultivars ◽  
Content Variable ◽  
Relationship Of ◽  
Olea Europaéa

AbstractThe olive tree (Olea europaeaL.) is one of the major cultivated species in the world, and Mediterranean countries produce about 90% of world cultivated olives. In this study, the genetic relationship of seven Turkish olive cultivars was investigated using non-codingtrnL-Fregions in chloroplastic genome. Cultivars demonstrated a similar sequence length of 330-340 bp with an average 35.26% G+C content. Variable (polymorphic/segregating), parsimony informative and total numbers of the insertion or the deletion of bases in the DNA (indel sites) were 4, 3, and 28, respectively. Nucleotide diversities π and θ were found as 0.00631 and 0.00644 respectively, while Tajima’s D was −0.786. cpDNAtrnL-Fregions of sequenced Turkish olive cultivars had a low level of genetic variations, and these non-coding regions were strictly conserved in all analyzed cultivars. Geographically distant shared more sequence similarities than relatively close cultivars. The phylogenetic analyses indicated that the biogeographic distribution of cultivars does not demonstrate any association inferring cultivar source. These results indicate the possibility of germplasm exchanges among countries or that some indel mutations contribute to variations of the Turkish olive gene pool. Thus, the authorities should develop the necessary programs to preserve the purity of native germplasms.

scholarly journals Development of a Competent and Trouble Free DNA Isolation Protocol for Downstream Genetic Analyses in Glycine Species

2016 ◽  
Vol 4 (8) ◽  
pp. 700 ◽  
Author(s):  
Muhammad Amjad Nawaz ◽  
Faheem Shehzad Baloch ◽  
Hafiz Mamoon Rehman ◽  
Bao Le ◽  
Fahima Akther ◽  
...  
Keyword(s):  
Molecular Biology ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Dna Isolation ◽  
Plant Molecular Biology ◽  
Pcr Amplification ◽  
Cost Effective ◽  
Extraction Methods ◽  
Glycine Species ◽  
Genomic Dna Isolation

Extraction of deoxyribose nucleic acid (DNA) from plants is preliminary step in molecular biology. Fast and cost effective genomic DNA isolation from Glycine species for downstream application is a major bottleneck. Here we report a high throughput and trouble free method for genomic DNA extraction from leaf and seeds of Glycine species with high quality and quantity. Protocol reports the optimization by employing different concentrations of CTAB and PVP in extraction buffer. Efficiency of optimized protocol was compared with frequently used DNA extraction methods. Wide adoptability and utility of this protocol was confirmed by DNA extraction from leaves as well as seeds of G. max, G. soja, G. tomentella and G. latifolia. Extracted DNA was successfully subjected to PCR amplification of five microsatellite markers and four putative glycosyltransferase genes. DNA extraction protocol is reproducible, trouble free, rapid and can be adopted for plant molecular biology applications.

scholarly journals Genetic relationship of penicillin resistant Streptococcus pneumoniae serotype 19B strains in Japan

1997 ◽  
Vol 118 (2) ◽  
pp. 105-110 ◽  
Author(s):  
R. YOSHIDA ◽  
Y. HIRAKATA ◽  
M. KAKU ◽  
H. TAKEMURA ◽  
H. TANAKA ◽  
...  
Keyword(s):  
Streptococcus Pneumoniae ◽  
Genetic Relationship ◽  
Gel Electrophoresis ◽  
Genomic Dna ◽  
Restriction Enzymes ◽  
Pulsed Field Gel Electrophoresis ◽  
Drug Resistant ◽  
Pulsed Field ◽  
Single Clone ◽  
Relationship Of

Pulsed field gel electrophoresis (PFGE) of the genomic DNA of penicillin resistant serotype 19B Streptococcus pneumoniae was carried out. Thirteen strains form the Nagasaki area and 12 strains from other areas in Japan were examined. Twenty-three strains were resistant to erythromycin, tetracycline and trimethoprim/sulfamethoxazole but susceptible to chloramphenicol. Eight strains were resistant to ceftriaxone. All strains were multiply resistant. Five strains isolated from Nagasaki were indistinguishable from each other by using restriction enzymes Apa I and Sma I. Two strains isolated from other areas were indistinguishable from the above five strains. We could classify 13 Nagasaki strains into 3 groups and the total of 25 Japanese strains into 6 groups. These results suggest that the increasing prevalence of multiply drug resistant S. pneumoniae serotyped 19B in Japan is not due to a single clone, but at least one clone has spread widely in Japan.

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