scholarly journals EFFICIENT METHOD OF DNA ISOLATION FROM BULKING SAMPLES OF SEEDLINGS

2021 ◽  
Vol 2021 (3) ◽  
pp. 29-48
Author(s):  
Irina Klimenko ◽  
Alexey Antonov ◽  
Vladimir Dushkin ◽  
Anastasia Shamustakimova ◽  
Yulian Mavlyutov
Keyword(s):  
Dna Isolation ◽  
Low Cost ◽  
Leaf Tissue ◽  
Perennial Grasses ◽  
Genetic Studies ◽  
Modified Method ◽  
Extraction Buffer ◽  
Low Efficiency ◽  
Commercial Kits ◽  
The Individual

Forage annual and perennial grasses are the difficult subject for molecular and genetic studies because of the problem with obtaining qualitative genomic DNA for PCR, due of high content of proteins, polysaccharides and polyphenols. The known methods of DNA extraction or the numerous commercial kits allow isolating purified nucleic acids from the leaf tissue, but characterized by low efficiency at seedlings using. The modified method of DNA isolation, based on the SDS-extraction buffer (sodium dodecil sulfate), is presented in this study. Significant modifications were introduced in the reagents compound and the steps of procedure accordingly to used type of plant tissue and the result was positive at usage on the bulking samples, as well as on the individual genotypes (the only seedling). Reliability of this method and the functionality of the obtained DNA samples were tested in PCR with different molecular markers (SSR, SRAP and PawS) in researches on revealing of forage legume grasses DNA polymorphism. The general advantages of the proposed method are simplicity and effectiveness, the possibility to isolate qualitative DNA without toxic reagents application, as well as relatively low cost and availability of reagents. This method can be useful for studying the genetic biodiversity and for decision the different tasks, required the rapid analysis of large plant populations.

scholarly journals Comparative analysis of the DNA isolated from thyme leaves using different methods

2020 ◽  
Vol 181 (3) ◽  
pp. 155-162
Author(s):  
I. V. Bulavin ◽  
O. A. Grebennikova ◽  
V. A. Brailko ◽  
S. A. Feskov ◽  
I. V. Mitrofanova
Keyword(s):  
Essential Oil ◽  
Cross Sections ◽  
Dna Isolation ◽  
Low Cost ◽  
Automated System ◽  
Ex Situ ◽  
Mass Analysis ◽  
High Quality ◽  
Cheap Method ◽  
Commercial Kits

Background. The base for a molecular analysis is DNA of high quality. For DNA isolation, different kits or classical methods are used. For mass analysis, isolation with kits is a very expensive process. So, the objective of our investigation was to find a cheap method for high-quality DNA isolation from leaves of various thyme cultivars.Materials and methods. Leaves cut from thyme accessions (Thymus mastichina L. cv. ‘Svetliachok’, T. striatus Vahl. cv. ‘Jubileiniy’, T. vulgaris L. cv. ‘Fantasia’, and T. vulgaris cv. ‘Jalos’.) maintained ex situ in the collection of the Nikita Botanical Gardens were used as the material for the analysis. Light microscopy was used to study leaf anatomy and localize essential oil on leaf cross sections. Essential oil was extracted on Ginsberg devices, and phenolic content was measured with The Folin–Ciocâlteu reagent (FCR). Commercial kits (DiamondDNATM, PureLink® Plant Total DNA Purification Kit) and classical methods (CTAB, CTAB with 2% polyvinylpyrrolidone) were used for DNA isolation. DNA quality was evaluated spectrophotometrically, with electrophoresis (horizontal, automated system Agilent 4200 TapeStation) and PCR.Results. The analysis showed that the leaf blade mesophyll of four thyme cultivars had inclusions with essential oil. The content of essential oil and phenolic compounds was measured biochemically. Since the plants were characterized by the presence of secondary metabolites, DNA was isolated by different methods. Spectrophotometry demonstrated that the classical CTAB method and CTAB with 2% PVP provided the best results. Using an automated electrophoresis system, the presence of high-molecularweight DNA (more than 52000 bp) in significant amounts was detected in the samples isolated with DiamondDNATM kit and CTAB + 2% PVP.Conclusion. Among the tested kits and methods, CTAB + 2% PVP provided thyme DNA suitable for PCR and, presumably, for genome library preparation. The low cost of reagents for this technique makes it applicable for future mass analysis of plant material. 

scholarly journals Comparison of methods for isolating fungal DNA

2012 ◽  
Vol 29 (Special Issue) ◽  
pp. S76-S85 ◽  
Author(s):  
P. Moťková ◽  
J. Vytřasová
Keyword(s):  
Dna Isolation ◽  
Dna Amplification ◽  
Food Samples ◽  
Plant Leaves ◽  
Pcr Inhibitors ◽  
Detection Techniques ◽  
Cell Wall Disruption ◽  
Fungal Dna ◽  
Commercial Kits ◽  
The Individual

In this study methods of fungal DNA isolation were optimised and compared. The aim of the isolation processes was to obtain DNA of sufficient quality and quantity necessary for its amplification, as most detection techniques require DNA amplification before the proper DNA detection itself. For this purpose, classic methods of DNA extraction were compared and optimised while isolations using commercial kits were also done. The methods were evaluated from several perspectives, with focus especially laid on the isolated DNA not contain PCR inhibitors which would prevent DNA amplification, thus inhibiting the detection itself. For optimising the individual methods, collection strains of the genus Aspergillus were used. After the evaluation, two most suitable methods were selected and chosen for isolating potentially aflatoxigenic moulds taken from food samples. These methods were the commercially supplied kit for isolating DNA from plant leaves from Sigma and a classic method according Cenis in combination with the cell wall disruption by means of liquid nitrogen.

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 Characterisation of a highly potent and near pan-neutralising anti-HIV monoclonal antibody expressed in tobacco plants

Retrovirology ◽  
2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Catherine M. Moore ◽  
Melanie Grandits ◽  
Clemens Grünwald-Gruber ◽  
Friedrich Altmann ◽  
Maria Kotouckova ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Low Cost ◽  
Leaf Tissue ◽  
Glycosylation Site ◽  
People Living With Hiv ◽  
Middle Income ◽  
Effector Cells ◽  
Extraction And Purification ◽  
Important Health ◽  
Different Strains

Abstract Background HIV remains one of the most important health issues worldwide, with almost 40 million people living with HIV. Although patients develop antibodies against the virus, its high mutation rate allows evasion of immune responses. Some patients, however, produce antibodies that are able to bind to, and neutralise different strains of HIV. One such ‘broadly neutralising’ antibody is ‘N6’. Identified in 2016, N6 can neutralise 98% of HIV-1 isolates with a median IC50 of 0.066 µg/mL. This neutralisation breadth makes N6 a very promising therapeutic candidate. Results N6 was expressed in a glycoengineered line of N. benthamiana plants (pN6) and compared to the mammalian cell-expressed equivalent (mN6). Expression at 49 mg/kg (fresh leaf tissue) was achieved in plants, although extraction and purification are more challenging than for most plant-expressed antibodies. N-glycoanalysis demonstrated the absence of xylosylation and a reduction in α(1,3)-fucosylation that are typically found in plant glycoproteins. The N6 light chain contains a potential N-glycosylation site, which was modified and displayed more α(1,3)-fucose than the heavy chain. The binding kinetics of pN6 and mN6, measured by surface plasmon resonance, were similar for HIV gp120. pN6 had a tenfold higher affinity for FcγRIIIa, which was reflected in an antibody-dependent cellular cytotoxicity assay, where pN6 induced a more potent response from effector cells than that of mN6. pN6 demonstrated the same potency and breadth of neutralisation as mN6, against a panel of HIV strains. Conclusions The successful expression of N6 in tobacco supports the prospect of developing a low-cost, low-tech production platform for a monoclonal antibody cocktail to control HIV in low-to middle income countries. Graphic abstract

A facile new modified method for the preparation of a new cerium-doped lanthanium cuperate perovskite energy storage system using nanotechnology

2021 ◽  
Author(s):  
Mervette El Batouti ◽  
H. A. Fetouh
Keyword(s):  
Solar Cells ◽  
Energy Storage ◽  
Dielectric Loss ◽  
Storage Systems ◽  
Low Cost ◽  
Storage System ◽  
Energy Storage System ◽  
Solar Ponds ◽  
Modified Method ◽  
Ferroelectric Perovskite

New ferroelectric perovskite sample: excellent dielectric, negligible dielectric loss for energy storage systems such as solar cells, solar ponds, and thermal collectors has been prepared at low cost using nanotechnology.

scholarly journals Potential uses of biomass from fast-growing crop miscanthus×giganteus

2012 ◽  
Vol 66 (2) ◽  
pp. 223-233 ◽  
Author(s):  
Nada Babovic ◽  
Gordana Drazic ◽  
Ana Djordjevic
Keyword(s):  
Panicum Virgatum ◽  
Large Scale ◽  
Low Cost ◽  
Water Concentration ◽  
High Water ◽  
Perennial Grasses ◽  
Great Promise ◽  
Cellulose Content ◽  
Production Chain ◽  
Miscanthus Giganteus

There is an increasing interest in perennial grasses as a renewable source of bioenergy and feedstock for second-generation cellulosic biofuels. Switchgrass (Panicum virgatum) and miscanthus (Miscanthus?giganteus), belonging to the parennial grasses group, are the major lignocellulosic materials being studied today as sources for direct energy production, biofuels, bioremediation and other. They have the ability to grow at low cost on marginal land where they will not compete with the traditional food crops. Miscanthus?giganteus possesses a number of advantages in comparison with the other potential energy crops such as are: high yields, low moisture content at harvest, high water and nitrogen use efficiencies, low need for annual agronomic inputs such as fertilizers and pesticides, high cellulose content, non-invasive character, low susceptibility to pests and diseases and broad adaptation to temperate growing environments. The main problems are low rate of survival during the first winter after the creation of plantation and the relatively high establishment costs. Miscanthus?giganteus is grown primarily for heat and electricity generation but can also be used to produce transport fuels. Miscanthus biomass has a very good combustion quality due to its low water concentration as well as its low Cl, K, N, S and ash concentrations compared to other lignocellulose plants. It is expected that miscanthus will provide cheaper and more sustainable source of cellulose for production of bioethanol than annual crops such as corn. Miscanthus has great promise as a renewable energy source, but it can only be realised when the grass production has been optimised for large-scale commercial cultivation. However, further research is still needed to optimise agronomy of miscanthus, to develop the production chain and pre-treatment as well as to optimise energy conversation route to produce heat, electricity, and/or fuels from biomass, if miscanthus is to compete with fossil fuel use and be widely produced.

scholarly journals ACCURACY ASSESSMENT OF UNDERWATER PHOTOGRAMMETRIC THREE DIMENSIONAL MODELLING FOR CORAL REEFS

2016 ◽  
Vol XLI-B5 ◽  
pp. 821-828 ◽  
Author(s):  
T. Guo ◽  
A. Capra ◽  
M. Troyer ◽  
A. Gruen ◽  
A. J. Brooks ◽  
...  
Keyword(s):  
Coral Cover ◽  
Accuracy Assessment ◽  
Low Cost ◽  
Three Dimensional ◽  
Point Clouds ◽  
3D Modelling ◽  
Relative Accuracy ◽  
3D Point Clouds ◽  
The Individual ◽  
Working Distance

Recent advances in automation of photogrammetric 3D modelling software packages have stimulated interest in reconstructing highly accurate 3D object geometry in unconventional environments such as underwater utilizing simple and low-cost camera systems. The accuracy of underwater 3D modelling is affected by more parameters than in single media cases. This study is part of a larger project on 3D measurements of temporal change of coral cover in tropical waters. It compares the accuracies of 3D point clouds generated by using images acquired from a system camera mounted in an underwater housing and the popular GoPro cameras respectively. A precisely measured calibration frame was placed in the target scene in order to provide accurate control information and also quantify the errors of the modelling procedure. In addition, several objects (cinder blocks) with various shapes were arranged in the air and underwater and 3D point clouds were generated by automated image matching. These were further used to examine the relative accuracy of the point cloud generation by comparing the point clouds of the individual objects with the objects measured by the system camera in air (the best possible values). Given a working distance of about 1.5 m, the GoPro camera can achieve a relative accuracy of 1.3 mm in air and 2.0 mm in water. The system camera achieved an accuracy of 1.8 mm in water, which meets our requirements for coral measurement in this system.

scholarly journals Development and Performance Testing of Solar Operated Insecticide and Pesticide Agro Spraying System

2019 ◽  
Vol 9 (1) ◽  
pp. 573-578
Keyword(s):  
Low Cost ◽  
Labour Productivity ◽  
Operating Time ◽  
Performance Testing ◽  
Maintenance Cost ◽  
Pump Efficiency ◽  
Agricultural Field ◽  
Low Efficiency ◽  
And Performance ◽  
Solar Powered

This paper represents the development and performance analysis of Solar operated Spraying system. Generally in the agricultural field, traditional conventional techniques like hand operated and fuel operated sprayer system for spraying pesticides have been used which is not eco-friendly, less labour productivity and low efficiency. These tools uses diesel as fuels which is harmful for the environment and also do increases the operating and maintenance cost. This motivates us to design and fabricate real-time product which is operated by solar energy. The main objective of this research is to design and fabricate the solar powered agricultural pesticide sprayer by considering parameters like desired spraying capacity, low weight, low cost, user-friendly nature, high operating time and for faster coverage of area. Mathematical models were developed after adopting suitable assumptions for calculation of power of the motor and sizing of battery, charge controller, solar panel required for spraying a known quantity of fluid. The parts required for the system had been selected by solving for known inputs values and considering their availability in the market. The maximum discharge at outlet of DC Pump, efficiency of pump had been calculated by taking different discharge at outlet of the pump. Further by using 12 Volt Led light, it can be operated in night mode and also is to reduce back pain of human being by keeping the tank in backside.

The Ethics of Conducting E-Mail Surveys

2004 ◽  
pp. 114-129 ◽  
Author(s):  
Sandeep Krishnamurthy
Keyword(s):  
Low Cost ◽  
Ethical Problem ◽  
Sampling Procedure ◽  
Academic Field ◽  
Mail Surveys ◽  
Effective Form ◽  
The Individual ◽  
Individual Contact ◽  
E Mail

E-mail is a low-cost and highly effective form of individual contact for primary research. However, researchers who contact strangers for their survey research through e-mail are, in essence, sending them Spam. Some academic researchers might argue that due to the low volume and infrequent nature of their surveys and the general positive perception of academia, their e-mail surveys do not add to the Spam problem. However, this is an insufficient resolution of the ethical problem. This chapter examines one solution to avoid this problem—the use of respondent permission prior to contact. Obtaining respondent permission is tricky and can be costly. But, it may be the only long-term solution. Importantly, using this approach could lead to a loss of randomness in the sampling procedure due to self-selection. Ideas for implementation of a permission-based contact system at the individual researcher and academic field level are provided at the end.

scholarly journals Evaluation of Extraction Buffers Using the Current Approach of Detecting Multiple Allergenic and Nonallergenic Proteins in Food

2004 ◽  
Vol 87 (6) ◽  
pp. 1458-1465 ◽  
Author(s):  
Carmen D Westphal ◽  
Marion R Pereira ◽  
Richard B Raybourne ◽  
Kristina M Williams
Keyword(s):  
Peanut Butter ◽  
Current Trend ◽  
Current Approach ◽  
Detection Methods ◽  
Food Allergens ◽  
Extraction Buffer ◽  
Sample Extraction ◽  
Working Together ◽  
Commercial Kits ◽  
Analytical Variation

Abstract The detection of food allergens has been a challenge because of the increasing need to ensure the absence of undeclared allergens in foods. The current trend in the detection of some food allergens, like peanuts, is based on the detection of multiple allergenic and nonallergenic proteins, and this is the approach that kit manufacturers have adopted. Because commercial kits differ in their ability to detect allergens, regulatory agencies, the food industry, and kit manufacturers are working together to standardize the detection methods. Three kits for the detection of peanuts have been evaluated for performance by the AOAC Research Institute. For this evaluation, a peanut butter suspension was used as a reference material. Several kit components contribute to between-kit analytical variation, even when the same sample is used. One component of commercial kits, which may be contributing to this variability, is the sample extraction buffer. In this study, differences in extractability of 3 allergenic foods were evaluated by using 4 different extraction buffers. The conclusion is that optimum allergen extractability was buffer-dependent, and no single buffer is appropriate for use as a universal extraction solution for all allergenic foods. Therefore, a thorough evaluation of sample preparation buffers needs to be performed for every individual allergenic food. In light of the results obtained, the current approach used for detection of peanut allergens based on the detection of multiple allergenic and nonallergenic proteins is being analyzed.

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