scholarly journals RANDOM AMPLIFIED POLYMORPHIC DNA (RAPDs) DETECTS PATTERNS OF GENETIC DIVERSITY IN DIPLOID MUSA ACUMINATA COLLA

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 660b-660
Author(s):  
Robert L. Jarret
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Geographic Origin ◽  
Morphological Characteristics ◽  
Principal Component ◽  
Chain Reaction ◽  
Banding Patterns ◽  
Random Primers ◽  
Pcr Products ◽  
Polymerase Chain

Patterns of diversity among thirty diploid clones of banana (Musa acuminata Colla.), collected in Papua New Guinea and the surrounding islands between 1987 and 1989, were examined genetically using the polymerase chain reaction (PCR) and random primers, to detect random amplified polymorphic DNA (RAPDs). PCR products were visualized on ethidium bromide stained agarose gels. Twenty of 60 random primers examined detected RAPDS in CTAB-extracted genomic DNA. Banding patterns ranged from very simple (1 or 2 bands/gel) to very complex (more than 20 bands/gel). All 30 Musa clones were distinguishable from each other based on their unique RAPD banding pattern. Principal component analysis (PCA) revealed several clusters of closely related clones within the materials examined. However, these clusterings were not correlated with either the geographic origin or the morphological characteristics of the clones. A role of the use of RAPDs in germplasm characterization is discussed.

scholarly journals Sources of Potential Errors in the Application of Random Amplified Polymorphic DNAs in Cucumber

HortScience ◽  
1996 ◽  
Vol 31 (2) ◽  
pp. 262-266 ◽  
Author(s):  
Jack Staub ◽  
Jeffery Bacher ◽  
Karl Poetter
Keyword(s):  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Inbred Lines ◽  
Sphaerotheca Fuliginea ◽  
Chain Reaction ◽  
Banding Patterns ◽  
Taq Dna Polymerase ◽  
Polymerase Chain ◽  
Stock Solutions ◽  
Pcr Conditions

The influence of tissue age, pathogen infestation, intrapopulation contamination, and polymerase chain reaction (PCR) conditions were assessed as sources of error in random amplified polymorphic DNA (RAPD) analysis. DNA from young, uninfected tissue provided the most consistent results. Plants infected with Sphaerotheca fuliginea Schl. (ex Fr.) Poll. showed variation in RAPD banding patterns compared to those of uninfected plants. Differences in banding patterns were detectable when DNA from two inbred lines were mixed at dilution ratios of ≤20:1 but not ≥50:1. Differing lots of commercially available 10× reaction buffer, MgCl2 stock solutions, and Taq DNA polymerase affected RAPD banding patterns and overall yield. For reproducibility of RAPD assays, it may be necessary to optimize reactions for specific lots of PCR reagents from either commercial or in-house sources.

scholarly journals Characterization of the Hyphal Swelling Group of Pythium: DNA Polymorphisms and Cultural and Morphological Characteristics

Plant Disease ◽  
1998 ◽  
Vol 82 (2) ◽  
pp. 218-222 ◽  
Author(s):  
K. Kageyama ◽  
H. Uchino ◽  
M. Hyakumachi
Keyword(s):  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Morphological Characteristics ◽  
Its Region ◽  
Restriction Enzymes ◽  
Dna Polymorphisms ◽  
Banding Patterns ◽  
Length Polymorphisms ◽  
Polymerase Chain

The hyphal swelling (HS) group of Pythium species and P. ultimum were studied for cultural and morphological characteristics, restriction fragment length polymorphisms of the amplified internal transcribed spacer (ITS) region in nuclear rDNA, and random amplified polymorphic DNA (RAPD) analysis of genome DNA. The shape of sporangia was spherical to subspherical or lemoniform and averaged 18.1–23.0 μm. All isolates could grow at 5 to 35°C, and the rate at the optimal temperature, 30°C, was 29–34 mm/24 h. The size of the ITS region amplified by polymerase chain reaction and the banding patterns after digestion with the restriction enzymes showed no variation between the HS group and P. ultimum. No difference in banding patterns was shown between the HS group and P. ultimum by RAPD analysis with each of three primers. Isolates examined were from Japan, and results should be confirmed from other regions.

scholarly journals Identifikasi Simplisia yang Dijual sebagai Strychnos ligustrina Bl. di Pasar Tradisional Surabaya dengan Metode Random Amplified Polymorphic DNA (RAPD)

2005 ◽  
Vol 11 (1) ◽  
pp. 19-24
Author(s):  
Oeke Yunita ◽  
Angelica Kresnamurti
Keyword(s):  
Cetyltrimethylammonium Bromide ◽  
Molecular Level ◽  
Random Amplified Polymorphic Dna ◽  
Local Market ◽  
Chain Reaction ◽  
Banding Patterns ◽  
Traditional Market ◽  
Ctab Method ◽  
Polymerase Chain ◽  
Rapd Method

Authentication of Strychnos ligustrina Bl. had been performed at molecular level (DNA) with Random Amplified Polymorphic DNA (RAPD) method, based on the amplification of random DNA fragments by Polymerase Chain Reaction (PCR) with a single arbitrary primer. The aim of this research was obtaining similar banding patterns between DNA of plant Strychnos ligustrina Bl. and DNA of its lignum on local market. Strychnos ligustrina Bl. was determined by UPT Balai Konservasi Tumbuhan Kebun Raya Purwodadi and plants sold as Strychnos ligustrina Bl. were collected as lignum from traditional market at Wonokromo, Rungkut, Genteng, Benowo dan Pabean. DNA from these plants were extracted by modified Cetyltrimethylammonium bromide (CTAB) method and amplified by RAPD method. Amplification had been performed by primer OPO-4 had shown banding patterns on the gel electrophoresis which banding patterns were shown by Strychnos ligustrina Bl. and plants sold as Strychnos ligustrina Bl. on Benowo. Based on this early result, we assume that plants sold as Strychnos ligustrina Bl. on Benowo has closely genetic relationship with Strychnos ligustrina Bl.

scholarly journals Detection and characterization of clustered regularly interspaced short palindromic repeat-associated endoribonuclease gene variants in Vibrio parahaemolyticus isolated from seafoods and environment

2019 ◽  
Vol 12 (5) ◽  
pp. 689-695
Author(s):  
Pallavi Baliga ◽  
Malathi Shekar ◽  
Moleyur Nagarajappa Venugopal
Keyword(s):  
Polymerase Chain Reaction ◽  
Vibrio Parahaemolyticus ◽  
Genotypic Variation ◽  
Gene Variants ◽  
Chain Reaction ◽  
Pcr Products ◽  
Short Palindromic Repeat ◽  
Polymerase Chain

Aim: In Vibrio parahaemolyticus, the clustered regularly interspaced short palindromic repeat (CRISPR)-associated cas6 endoribonuclease gene has been shown to exhibit sequence diversity and has been subtyped into four major types based on its length and composition. In this study, we aimed to detect and characterize the cas6 gene variants prevalent among V. parahaemolyticus strains isolated from seafoods and environment. Materials and Methods: Novel primers were designed for each of the cas6 subtypes to validate their identification in V. parahaemolyticus by polymerase chain reaction (PCR). In total, 38 V. parahaemolyticus strains isolated from seafoods and environment were screened for the presence of cas6 gene. Few representative PCR products were sequenced, and their phylogenetic relationship was established to available cas6 gene sequences in GenBank database. Results: Of the 38 V. parahaemolyticus isolates screened, only about 40% of strains harbored the cas6 endoribonuclease gene, among which 31.6% and 7.9% of the isolates were positive for the presence of the cas6-a and cas6-d subtypes of the gene, respectively. The subtypes cas6-b and cas6-c were absent in strains studied. Sequence and phylogenetic analysis also established the cas6 sequences in this study to match GenBank sequences for cas6-a and cas6-d subtypes. Conclusion: In V. parahaemolyticus, the Cas6 endoribonuclease is an associated protein of the CRISPR-cas system. CRISPR-positive strains exhibited genotypic variation for this gene. Primers designed in this study would aid in identifying the cas6 genotype and understanding the role of these genotypes in the CRISPR-cas immune system of the pathogen.

scholarly journals Heavy metal-induced oxidative stress and DNA damage as shown by RAPD-PCR in leaves of Elodea canadensis

2020 ◽  
Vol 3 (1) ◽  
pp. 14
Author(s):  
Dilek Demirezen Yilmaz ◽  
Nuri Ercan ◽  
Fahriye Sumer Ercan
Keyword(s):  
Heavy Metals ◽  
Dna Damage ◽  
Random Amplified Polymorphic Dna ◽  
Antioxidant Response ◽  
Elodea Canadensis ◽  
Chain Reaction ◽  
Banding Patterns ◽  
Dna Damages ◽  
Rapd Pcr ◽  
Polymerase Chain

The objective of the study was to evaluate the antioxidant response and DNA damage of heavy metals (Cd, Cu and Cr) in Elodea canadensis. Superoxide dismutase (SOD), catalase (CAT), Glutathione reductase (GR) and lipid peroxidation levels of leaves of Elodea canadensis which was exposed to different concentrations of heavy metals (Cd: 2, 5, 10, 15, 25 ppb; Cu: 200, 500, 1000, 2500 ppb and Cr: 1, 3, 10, 15, 25 ppb) in a hydroponic culture were determined spectrophotometrically. The highest induction in SOD and CAT activities were determined at highest concentration of heavy metals. The Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) technique was used to investigate the variation of DNA banding patterns of samples that exposed to different concentrations of heavy metals. Changing in band intensity and the gain and loss of bands were demonstrated the genotoxic effect of heavy metals. Bioaccumulation, oxidative responses and DNA damages were shown that Elodea canadensis represents a useful bioindicator.

scholarly journals AuNPs-Based Colorimetric Assay for Identification of Chicken Tissues in Meat and Meat Products

2015 ◽  
Vol 2015 ◽  
pp. 1-6
Author(s):  
Hejing Han ◽  
Wen Yi ◽  
Dongjun Hou ◽  
Tingting Huang ◽  
Zhihui Hao
Keyword(s):  
Target Gene ◽  
Colorimetric Assay ◽  
Meat Products ◽  
Chain Reaction ◽  
Chicken Tissues ◽  
Pcr Products ◽  
Gene Polymerase Chain Reaction ◽  
D Loop ◽  
Polymerase Chain

A simple colorimetric assay was developed to identify chicken tissues in meat and meat products by utilizing thiol-labeled primers and unmodified gold nanoparticles (AuNPs). Primers were designed based on the chicken-specific mitochondrial D-loop gene. Polymerase chain reaction (PCR) is applied to amplify the target gene, and the PCR products labeled with thiol at one end were obtained. Following the mixing of AuNPs with the PCR products, the thiol binds to the surface of AuNPs, resulting in the formation of GNP-PCR products. The resultant PCR products had abundant negative charges, which made AuNPs maintain dispersion under the role of electrostatic repulsion. As a result, in the presence of PCR products, AuNPs remained red in the presence of salt. In the absence of PCR products, the color of AuNPs changed from red to blue; therefore, the method described here could be exploited for the verification of chicken tissues with high accuracy.

scholarly journals Universally primed polymerase chain reaction analysis of Fusarium avenaceum isolated from wheat and barley in Finland

1997 ◽  
Vol 6 (1) ◽  
pp. 25-36 ◽  
Author(s):  
Tapani Yli-Mattila ◽  
Nina V. Mironenko ◽  
Irina A. Alekhina ◽  
Asko Hannukkala ◽  
Sergey A. Bulat
Keyword(s):  
Phylogenetic Analyses ◽  
Geographic Origin ◽  
Polymerase Chain Reaction Analysis ◽  
Fusarium Avenaceum ◽  
Universal Primers ◽  
Clear Correlation ◽  
Chain Reaction ◽  
Pcr Products ◽  
Pathogenicity Tests ◽  
Polymerase Chain

Twenty-two Fusarium avenaceum isolates from Finnish wheat and barley were analysed using the chain reaction with universal primers (UP-PCR). Each isolate could be distinguished from others by UP-PCR products on polyacrylamide gels. The isolates tested were clustered into two main groups and further into several subgroups by UP-PCR profiles and phylogenetic analyses. The phylogenetic relationships of these groups are discussed. No clear correlation was found between the groups and host plant preference or the geographic origin of F. avenaceum isolates. Pathogenicity tests showed differences between F. avenaceum isolates, but two isolates, one from wheat and the other from barley, were the most aggressive in wheat and barley. This fungus, usually known as a weak pathogen of cereals and other crops, has thus probably not evolved in respect to its ability to damage wheat or barley.

scholarly journals Optimasi Konsentrasi DNA dan MgCl2 pada Reaksi Polymerase Chain Reaction-Random Amplified Polymorphic DNA untuk Analisis Keragaman Genetik Tanaman Faloak (Sterculia quadrifida R.Br) (Optimization of DNA and MgCl2 Concentrations in Polymerase Chain Reacti

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Uslan Uslan ◽  
Made Pharmawati
Keyword(s):  
Genetic Diversity ◽  
Polymerase Chain Reaction ◽  
Random Amplified Polymorphic Dna ◽  
Optimum Conditions ◽  
Chain Reaction ◽  
Dna Templates ◽  
Clear Band ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Pcr Conditions

Abstrak Faloak  merupakan tanaman yang tumbuh di lahan kritis. Sebagai upaya mendukung pemuliaan dan konservasi tanaman faloak diperlukan informasi keragaman genetiknya. Salah satu metode analisis keragaman genetik adalah menggunakan penanda DNA yang berbasis PCR. Untuk itu diperlukan kondisi PCR (Polymerase Chain Reaction) yang tepat sehingga diperoleh hasil yang dapat dianalisis lebih lanjut. Penelitian ini bertujuan menentukan kondisi optimum PCR-RAPD (Polymerase Chain Reaction-Random Amplified Polymorphic DNA) tanaman faloak. Ekstraksi DNA dilakukan dengan metode CTAB. Optimasi dilakukan dengan menggunakan beberapa konsentrasi DNA cetakan dan MgCl2. Kondisi optimum PCR-RAPD tanaman faloak yang menghasilkan pita produk PCR yang jelas diperoleh  menggunakan 50 ng/ul DNA, 3 mM MgCl2 serta jumlah siklus termal 45 x. Kata kunci : PCR-RAPD, optimasi, tanaman faloak Abstract Faloak is a plant that grows on critical lands. In an effort to support breeding and conservation of faloak, information about its genetic diversity is required. One of the methods of genetic diversity analysis is using PCR-based DNA markers. For that purpose, proper PCR conditions is needed in order to obtain results that can be further analyzed. This study aimed to determine the optimum conditions for PCR-RAPD of faloak plants. DNA extraction was conducted using CTAB. Optimization was done by using several concentrations of DNA templates and MgCl2. The optimum conditions of PCR-RAPD of faloak plants that produce clear band of PCR products were obtained using 50 ng/ ul DNA, 3 mM MgCl2 and 45x thermal cycles Keywords : PCR-RAPD, optimization, faloak plant

scholarly journals A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Dna Markers ◽  
5S Rdna ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Species Specific ◽  
Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

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