scholarly journals Evaluation of eight protocols for genomic DNA extraction of Hypostomus commersoni Valenciennes, 1836 (Loricariidae: Siluriformes)

2019 ◽  
Author(s):  
P. Mezzomo ◽  
A. A. Mielniczki-Pereira ◽  
T. L. Sausen ◽  
J. R. Marinho ◽  
R. L. Cansian
Keyword(s):  
Dna Extraction ◽  
Genetic Material ◽  
Proteinase K ◽  
Ocular Tissue ◽  
South American ◽  
Target Tissue ◽  
Sample Collection ◽  
High Concentration ◽  
Positive Spectrum

AbstractThe principle and the techniques applied in DNA extraction play a pivotal role in the obtention of a purified genetic material. The present study investigates the efficiency of eight protocols in the DNA extraction of Hypostomus commersoni, an essential component of South American freshwater ichthyofauna. The quality of samples was assessed through spectrophotometry, gel electrophoresis, and PCR-RAPD amplification. The efficiency of DNA extraction was influenced both by the method applied and the target-tissue of choice. Higher concentrations and yield of DNA were obtained from ocular tissue, with a positive spectrum of incubation in lysis buffer for up to 36 hours after sample collection, using fresh tissues and in the presence of a high concentration of Proteinase K (20 mg.ml-1). In these conditions, samples were successfully amplified. To date, there is no record of description for the parameters analyzed in this work, neither the description of RAPD markers for the species H. commersoni.

scholarly journals Evaluation of eight protocols for genomic DNA extraction of Hypostomus commersoni Valenciennes, 1836 (Loricariidae: Siluriformes)

2020 ◽  
Author(s):  
P. Mezzomo ◽  
A. A. Mielniczki-Pereira ◽  
T. L. Sausen ◽  
J. R. Marinho ◽  
R. L. Cansian
Keyword(s):  
Dna Extraction ◽  
Rapd Markers ◽  
Genetic Material ◽  
Proteinase K ◽  
Ocular Tissue ◽  
South American ◽  
Target Tissue ◽  
Sample Collection ◽  
High Concentration ◽  
Positive Spectrum

Abstract The principle and the techniques applied in DNA extraction play a pivotal role in the obtention of a purified genetic material. The present study investigates the efficiency of eight protocols in the DNA extraction of Hypostomus commersoni, an essential component of South American freshwater ichthyofauna. The quality of samples was assessed through spectrophotometry, gel electrophoresis, and PCR-RAPD markers amplification. The efficiency of DNA extraction was influenced both by the method applied and the target-tissue of choice. Higher concentrations and yield of DNA were obtained from ocular tissue, with a positive spectrum of incubation in lysis buffer for up to 36 hours after sample collection, using fresh tissues and in the presence of a high concentration of Proteinase K (20 mg.ml-1). In these conditions, samples were successfully amplified. To date, there is no record of description for the parameters analyzed in this work, neither the description of RAPD markers for the species H. commersoni.

scholarly journals Comparison of DNA extraction using proteinase K and extraction kit: analysis of the quality of the genetic material

2018 ◽  
Vol 54 (2) ◽  
Author(s):  
André Luís F. Santos ◽  
Carolina Q. P. Oliveira ◽  
Geovana Nicole P. N. Arruda ◽  
John Kenned Martins
Keyword(s):  
Dna Extraction ◽  
Genetic Material ◽  
Proteinase K

scholarly journals UNA METODOLOGÍA NO INVASIVA PARA LA EXTRACCIÓN DE ADN DE CORYPHAENA HIPPURUS

2016 ◽  
Vol 44 (2) ◽  
Author(s):  
Bruna Rafaela Caetano Nunes Pazdiora ◽  
John Josephraj Selvaraj ◽  
Diego Fernando Marmolejo ◽  
Jaime Eduardo Muñoz ◽  
Ángela Inés Guzmán
Keyword(s):  
Dna Extraction ◽  
Economic Value ◽  
Animal Tissue ◽  
Proteinase K ◽  
Sample Collection ◽  
Non Invasive ◽  
Coryphaena Hippurus ◽  
Molecular Studies ◽  
Invasive Method ◽  
Tissue Material

A protocol for DNA extraction and effect of the type of tissue material for obtaining DNA from dolphinfih in high quantity and quality for molecular studies was evaluated. The DNA of 25 individuals was extracted from the digestion of the muscle and the dorsal fi, with proteinase K, and the extraction kit for blood and animal tissue. DNA were obtained in all the samples; however, dorsal fis samples showed signifiantly higher amount of DNA than muscle samples. The defiition of an appropriate methodology of DNA extraction can contribute to studies of genetic structure of the species. DNA extraction from dolphinfih dorsal fis, non-invasive method, is an alternative that is environmentally, socially, and commercially viable, where sample collection does not require the sacrifie of the animal and do not alter its economic value.

scholarly journals Genetic diversity and comparative study of genomic DNA extraction protocols in Tamarix L. species

Caryologia ◽  
2021 ◽  
Vol 74 (2) ◽  
pp. 131-139
Author(s):  
Xiao Cheng ◽  
Xiaoling Hong ◽  
Majid Khayatnezhad ◽  
Fazal Ullah
Keyword(s):  
Genetic Diversity ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Species Group ◽  
Proteinase K ◽  
Amplification Efficiency ◽  
Nrdna Its ◽  
Dna Extraction Method ◽  
Dna Contents

The genus Tamarix  consists of about 54 species that mainly grow in saline areas of deserts and semi-deserts. This genus is chemically characterized by the presence of tannins, flavonoids, anthocyanins and essential oils which interfere with the extraction of pure genomic DNA. Thus it is necessary to optimize extraction protocols to minimize the influence of these compounds to the lowest level. The present study compares the efficiency of five different approaches to extract total genomic DNA in Tamarix species, showing significant differences in the extracted DNA contents and quality,by using  Kit (DNP TM Kit), CTAB DNA extraction method by Murray and Thompson, Sahu et al., Nalini et al. and Bi et al., for the extraction of DNA from Tamarix species. Our results showed significant differences in DNA contents between these five methods. The quantity and quality of extracted genomic DNA were checked by the spectrophotometer, Nano-Drop and and agarose gel electrophoresis analysis. Finally, a PCR-based method was also applied to verify the amplification efficiency for two molecular markers (ITS and ISSR).. In the present study, the genetic diversity of 96 Tamarix individuals species and 8 populations were studied using 10 ISSR markerswhile for nrDNA ITS 8 species samples were used. The method of Nalini et al., provided best results (207 ng/μL) in terms of quantity and quality ofDNA. Our results proposed that this method could be effective for plants with the same polysaccharides, proteins and polyphenols components. The advantage of this method is simple and fast as it does not involve time consuming steps such as incubation at higher temperatures, and also do not requires expensive chemicals such as proteinase K, liquid nitrogen. ,. The success of this method in obtaining high-quality genomic DNA has been demonstrated in the Tamarix species group and the reliability of this method has been discussed.

AN IMPROVED METHOD FOR INHIBITOR FREE NUCLEIC ACIDS FROM POLYPHENOL RICH LEAVES OF MUSA SPP

2017 ◽  
Vol 23 (1) ◽  
Author(s):  
N.NANDHA KUMAR ◽  
K. SOURIANATHA SUNDARAM ◽  
D. SUDHAKAR ◽  
K.K. KUMAR
Keyword(s):  
Polymerase Chain Reaction ◽  
Quantitative Polymerase Chain Reaction ◽  
Proteinase K ◽  
Chain Reaction ◽  
Banana Plant ◽  
High Molecular Weight Dna ◽  
Musa Spp ◽  
Leaf Tissues ◽  
Polymerase Chain

Excessive presence of polysaccharides, polyphenol and secondary metabolites in banana plant affects the quality of DNA and it leads to difficult in isolating good quality of DNA. An optimized modified CTAB protocol for the isolation of high quality and quantity of DNA obtained from banana leaf tissues has been developed. In this protocol a slight increased salt (NaCl) concentration (2.0M) was used in the extraction buffer. Polyvinylpyrrolidone (PVP) and Octanol were used for the removal of polyphenols and polymerase chain reaction (PCR) inhibitors. Proteins like various enzymes were degraded by Proteinase K and removed by centrifugation from plant extract during the isolation process resulting in pure genomic DNA, ready to use in downstream applications including PCR, quantitative polymerase chain reaction (qPCR), ligation, restriction and sequencing. This protocol yielded a high molecular weight DNA isolated from polyphenols rich leaves of Musa spp which was free from contamination and colour. The average yields of total DNA from leaf ranged from 917.4 to 1860.9 ng/ìL. This modified CTAB protocol reported here is less time consuming 4-5h, reproducible and can be used for a broad spectrum of plant species which have polyphenol and polysaccharide compounds.

Ground water quality of selected areas of Punjab and Sind Provinces, Pakistan: Chemical and microbiological aspects

2019 ◽  
Author(s):  
Chem Int
Keyword(s):  
Ionic Concentration ◽  
Chloride Ions ◽  
Potassium Ions ◽  
Ammonium Ions ◽  
High Concentration ◽  
Phosphate Ions ◽  
Sodium Magnesium ◽  
Very High ◽  
Microbiological Aspects

The assessment of groundwater is essential for the estimation of suitability of water for safe use. An attempt has been made to study the groundwater of selected areas of Punjab (Sheikhupura & Sahiwal) and Sindh (Sindh, Jawar Dharki and Dharki), Pakistan. The results indicate that pH, color and odor were all within limits of WHO that is pH ranges 6.5–8.5, colorless and odorless, respectively. The high values of suspended solids were observed in the Sindh-1 and Dharki samples. Microbiologically only Sahiwal and Jawar Dharki were found fit for drinking purpose. Trace metals analysis of Sheikhupura-1 and Sindh-1 showed that values do not fall within limits of WHO for Iron. The ionic concentration analysis showed that high bicarbonate (HCO3-), ions are present in the samples of Sahiwal and Dharki; Sindh-1 and Jawar Dharki samples showed very high concentration for chloride ions, all samples were satisfactory level for sulphate (SO42-), sodium, magnesium and phosphate ions except samples of Sindh-1 and Jawar Dharki. High concentration of calcium and potassium ions was observed in samples of Sindh-1, while all other samples were found fit for drinking purposes in respect of nitrate, nitrite and ammonium ions. The high concentration of Fluoride was found only in Sheikhupura-2 samples.

Quality Improvement of the DNA extracted by boiling method in Gram negative bacteria

2017 ◽  
Vol 6 (04) ◽  
pp. 5347 ◽  
Author(s):  
Omar B. Ahmed* ◽  
Anas S. Dablool
Keyword(s):  
Dna Extraction ◽  
Control Method ◽  
Extraction Procedure ◽  
Cost Effective ◽  
High Yield ◽  
Gram Negative Bacteria ◽  
Bacterial Dna ◽  
Gram Negative ◽  
E Coli

Several methods of Deoxyribonucleic acid (DNA) extraction have been applied to extract bacterial DNA. The amount and the quality of the DNA obtained for each one of those methods are variable. The study aimed to evaluate bacterial DNA extraction using conventional boiling method followed by alcohol precipitation. DNA extraction from Gram negative bacilli was extracted and precipitated using boiling method with further precipitation by ethanol. The extraction procedure performed using the boiling method resulted in high DNA yields for both E. coli and K. pneumoniae bacteria in (199.7 and 285.7μg/ml, respectively) which was close to control method (229.3 and 440.3μg/ml). It was concluded that after alcohol precipitation boiling procedure was easy, cost-effective, and applicable for high-yield quality of DNA in Gram-negative bacteria.

scholarly journals Methodological Approaches to DNA Authentication of Foods, Wines and Raw Materials for Their Production

Foods ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 595
Author(s):  
Aram G. Galstyan ◽  
Vladislav K. Semipyatniy ◽  
Irina Yu. Mikhailova ◽  
Khamid Kh. Gilmanov ◽  
Alana V. Bigaeva ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Dna Extraction ◽  
Raw Materials ◽  
Direct Sequencing ◽  
Genetic Identification ◽  
Proteinase K ◽  
Vitis Vinifera L ◽  
Acid Extraction ◽  
Plant Component ◽  
Sensitivity Specificity

DNA authentication of wines is a process of verifying their authenticity by genetic identification of the main plant component. The sample preparation of experimental and commercial wines was carried out by precipitation of wine debris by centrifugation with preliminary exposure with precipitators and co-precipitators, including developed macro- and micro- volume methods applicable to white or red wines, using polyvinylpyrrolidone as a co-precipitator. Addition of 2-mercaptoethanol and proteinase K to the lysing solution made it possible to adapt the technology for DNA extraction from the precipitated wine debris. The additionally tested technique of DNA extraction from wine debris by dimethyl sulfoxide (DMSO) lysis had fewer stages and, consequently, a lower risk of contamination. The results of further testing of one of the designed primer pairs (UFGT-F1 and UFGT-R1) in conjunction with the tested methods of wine material sample preparation and nucleic acid extraction, showed the advantage in the given set of oligonucleotides over previously used ones in terms of sensitivity, specificity and reproducibility. The developing strategy for genetic identification of grape varieties and DNA authentication of wines produced from them based on direct sequencing of polymerase chain reaction (PCR) products is implemented by interpreting the detected polymorphic positions of variable Vitis vinifera L. UFGT gene locus with distribution and split into 13 UFGT gene-associated groups.

Arguments for ‘ocular donation’ as standardised terminology to reduce the ‘ick factor’ of ‘eye donation’

2022 ◽  
pp. medethics-2021-108003
Author(s):  
Katrina A Bramstedt
Keyword(s):  
Life Support ◽  
Deceased Donor ◽  
Informed Consent Process ◽  
Tissue Donation ◽  
Ocular Tissue ◽  
Corneal Tissue ◽  
Root Cause ◽  
Eye Donation ◽  
Over Time

This brief report presents the global problem of the shortfall of donor corneal tissue for transplantation, a potential root cause (‘ick factor’ language), and a potential solution (modification of ‘ick factor’ language). Specifically, use of the term ‘eye donation’ is a potential hurdle to ocular tissue donation as it can stimulate the ‘ick factor.’ Verbiage such as ‘ocular (eye tissue)’ could be a method of providing terminology that is less emotive than ‘eye donor’ or ‘eye donation.’ The field of transplantation has experienced terminology shifts over time; for example, ‘cadaver’ has been replaced with ‘deceased donor,’ ‘harvest’ has been replaced with ‘recover,’ and ‘life support’ has been replaced with ‘ventilated.’ Notably, only a small number of regions worldwide are using ‘ocular’ terminology, yet it could be an important step to enhancing the informed consent process and improving donation rates, potentially increasing transplant and optimising patient quality of life for those with treatable blindness.

Potential strategies for cancer gene therapy

2021 ◽  
Author(s):  
Moataz Dowaidar
Keyword(s):  
Gene Therapy ◽  
Clinical Stage ◽  
Genetic Material ◽  
Direct Methods ◽  
Cancer Gene Therapy ◽  
Target Cells ◽  
Target Tissue ◽  
Cancer Gene ◽  
P Gene

Gene therapy involves transferring genetic material (DNA or RNA) to repair, regulate or replace genes to cure a disease. One of the most crucial barriers is successful delivery of the targeted gene into the target tissue. Various vector-based approaches have been developed to deliver the transgene to the target cells. In different cancers, numerous of these vectors are being developed for purposes such as immunotherapy, suicide gene therapy, microRNA (miRNA) focused treatment, oncogene silencing, and gene editing using CRISPR/Cas9. This article reviews several alternatives to cancer gene therapy, as well as their preclinical and clinical outcomes, possible limitations, and overall therapy effects. Ways of delivering cancer gene therapy include direct methods for introducing genetic material. Nonviral vectors are easy to manufacture and may be chemically modified to increase their usefulness. Cationic polymers such as Poly-L-Lysine (PLL) and Polyethylenimine (PEI-SS) are the most extensively used polycationic polymers for gene transfer, particularly in vitro. Many RNAi-based therapeutic approaches are approaching the clinical stage, and nanocarriers are likely to play a crucial role in treating specific cancers. In the previous decade, non-viral approaches were used in more than 17 percent of all gene therapy trials. The message is that this is a safe and effective technique for transferring genes to cancer patients who need it to be a safe, successful therapy. Exosomes were developed to carry oncogene-specific short interfering RNA. Sushrut and colleagues revealed that exosomes provide superior carriers of short RNA and prevent tumor growth than liposomes. Inhalation-based gene therapy (aerosol-mediated gene delivery) has gained pace as a feasible treatment approach, especially for lung cancer. Because the intended transgene is steered to specific cells/tissues, this should further increase therapeutic efficiency.

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