scholarly journals Effect of Flaxseed on some hormonal profile and genomic DNA concentration in Karadi lambs

2019 ◽  
Vol 388 ◽  
pp. 012035 ◽  
Author(s):  
A N Yousif
Keyword(s):  
Genomic Dna ◽  
Hormonal Profile ◽  
Dna Concentration

scholarly journals Optimization of DNA concentration to amplify short tandem repeats of human genomic DNA

2012 ◽  
Vol 12 (4) ◽  
pp. 236 ◽  
Author(s):  
Fahri Q. Gavazaj ◽  
Ilia I. Mikerezi ◽  
Valon H. Morina ◽  
Fatmir A. Cakaj ◽  
Ekrem B. Maloku ◽  
...  
Keyword(s):  
Genomic Dna ◽  
Short Tandem Repeats ◽  
Tandem Repeats ◽  
Dna Concentration ◽  
Human Genomic ◽  
Human Genomic Dna ◽  
Short Tandem

scholarly journals Preservation and maceration of amazon açai leaflet tissue to obtain genomic DNA

2019 ◽  
Vol 35 (4) ◽  
Author(s):  
Hellen Sandra Freires da Silva Azêvedo ◽  
Polinar Bandeira Rufino ◽  
José Marlo Araújo de Azevedo ◽  
Luciélio Manoel da Silva ◽  
Lúcia Helena de Oliveira Wadt ◽  
...  
Keyword(s):  
Liquid Nitrogen ◽  
Silica Gel ◽  
Genomic Dna ◽  
Leaf Tissue ◽  
Dna Concentration ◽  
Randomized Design ◽  
Molecular Studies ◽  
Tissue Maceration ◽  
Genomic Regions ◽  
Transport Buffer

The objective of this study was to test the efficiency of preservation and maceration methods for Euterpe precatoria leaflet tissue to obtain genomic DNA for molecular studies. The leaflets of E. precatoria were collected in an experimental field at Embrapa Acre, Brazil. The study was conducted in a completely randomized design with 10 replicates, in a 12 × 2 factorial structure, with 12 storage treatments (fresh; lyophiliser 3 days; refrigerator 3, 5, and 7 days; silica gel 7, 10, 20, and 30 days; and transport buffer 3, 5, and 7 days) and two leaf tissue maceration methods (liquid nitrogen and the TissueLyser®). Statistically significant differences in the obtained DNA concentration were found between the maceration and storage treatments. The TissueLyser® macerator produced higher DNA concentrations when compared to liquid nitrogen. For the storage treatments, five groups were formed based on DNA concentration when macerated with the TissueLyser® and two groups when macerated with liquid nitrogen. The DNA concentrations ranged from 285.00 ng/µL (7 days in transport buffer) to 702.00 ng/µL (30 days in silica gel) when the leaflets were macerated with liquid nitrogen, and from 572.73 ng/µL (30 days in silica gel) to 2,850.00 ng/µL (3 days in lyophiliser) using the TissueLyser® macerator. The DNA purity (A260/A280 nm) varied from 1.30 to 1.70 when the leaflets were macerated with liquid nitrogen and from 1.30 to 1.90 with the TissueLyser® macerator. Despite the variations in leaf tissue preservation and DNA concentration, all treatments were effective for DNA isolation and it was possible to amplify genomic regions of microsatellite markers by PCR. It was concluded that leaflets of E. precatoria stored in a lyophiliser and processed with an automatic macerator resulted in satisfactory DNA for molecular studies.

scholarly journals Effect of Proteinase-K on Genomic DNA Extraction from Gram-positive Strains

1970 ◽  
Vol 4 (1) ◽  
pp. 53-57 ◽  
Author(s):  
Mohammad Shahriar ◽  
Md Rashidul Haque ◽  
Shaila Kabir ◽  
Irin Dewan ◽  
Mohiuddin Ahmed Bhuyian
Keyword(s):  
Bacillus Subtilis ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Procedure ◽  
Proteinase K ◽  
Clinical Samples ◽  
Gram Positive ◽  
Genomic Dna Extraction ◽  
Dna Concentration ◽  
The Mean

Direct extraction of DNA from natural environment and clinical samples has become a useful alternative for the phylogenetic identification and in situ detection of individual microbial cells without cultivation. In this study, three different Gram positive microorganisms (B. cereus, B. subtilis, and S. aureus) were chosen for genomic DNA extraction. High salt SDS (Sodium Dodesyl Sulfate) based extraction method was followed to extract genomic DNA with addition of three different lysis protocols to observe the effect of proteinase-K on total genomic DNA yield, lysis steps were carried with SDS, SDS with 3 μl proteinase-K and SDS with 6μl proteinase-K. High molecular weight intact DNA bands were observed only for Bacillus subtilis when the extraction procedure was carried out in presence of SDS, SDS with proteinase-K (3μl) and SDS with increased amount of proteinase-K (6μl). In presence of SDS and increased amount of proteinase-K (6μl) the mean value of DNA concentration for Bacillus cereus, Bacillus subtilis, and Staphylococcus aureus were found to be 1.53±0.15, 1.36±0.10 and 1.65±0.10 μg/μl respectively. However, in absence of proteinase-K, the mean values of DNA concentration were found to be decreased (1.28±0.10, 1.34±0.15, 1.23±0.10 μg/μl for B. cereus, B. subtilis, and S. aureus respectively) for all these stains. Although in case of B. subtilis the overall effect of proteinase-K was not found to be significant in terms of DNA concentration and DNA band intensity, however, for B. cereus, and S. aureus sharp decrease in total extracted DNA concentration was observed suggesting the increased lysis effect of proteinase-K on the thick peptidoglycan layer of Gram-positive cell wall such as B. cereus, and S. aureus.   Key words: Extraction; Genomic DNA; Lysis buffer; Gram positive organism. DOI: http://dx.doi.org/10.3329/sjps.v4i1.8867 SJPS 2011; 4(1): 53-57

scholarly journals Plant Genomic DNA Extraction for Selected Herbs and Sequencing their Internal Transcribed Spacer Regions Amplified by Specific Primers

2016 ◽  
Vol 11 (10) ◽  
pp. 1934578X1601101
Author(s):  
Farah Izana Abdullah ◽  
Lee Suan Chua ◽  
Zaidah Rahmat ◽  
Azman Abd Samad ◽  
Alina Wagiran
Keyword(s):  
Dna Extraction ◽  
Plant Species ◽  
Dna Sequences ◽  
Incubation Time ◽  
Internal Transcribed Spacer ◽  
Genomic Dna ◽  
Molecular Technique ◽  
Hexadecyltrimethylammonium Bromide ◽  
Genomic Dna Extraction ◽  
Dna Concentration

This study was focused on plant genomic DNA extraction and sequencing from five commonly used medicinal herbs, namely Impatiens balsamina, Ficus deltoidea, Centella asiatica, Andrographis paniculata and Orthosiphon aristatus. This molecular technique is another highly reliable alternative for plant species identification besides phytochemical profiling. Three cetyl hexadecyltrimethylammonium bromide (CTAB) based methods with slight modification on incubation time, salt content and other additives were used for DNA extraction. The CTAB method of Doyle and Doyle produced higher DNA concentration from I. balsamina, most probably due to the presence of ammonium acetate in the washing buffer and longer incubation time (2 h). The CTAB based method was suitable for A. paniculata because a high DNA concentration of acceptable quality was obtained for all the modified methods. However, O. aristatus was likely to have a lower DNA concentration (33–87 μg/g) and quality, probably due to the high concentration of phenolic compounds, particularly rosmarinic acid. The extracted genomic DNA was effectively amplified by a polymerase chain reaction using a universal primer of internal transcribed spacer (ITS), particularly AB101 and AB102 at the optimum annealing temperature of 48°C. The DNA sequences were analyzed by phenetic analysis and it was found that they have high similarity with the nucleotide sequences of ITS regions for similar plant species in the GenBank database of the National Center for Biotechnology Information.

Isopropanol DNA Precipitation - Best for DNA Concentration v1

2021 ◽  
Author(s):  
Lynn Doran
Keyword(s):  
Aqueous Solution ◽  
Molecular Weight ◽  
New England ◽  
Genomic Dna ◽  
Dna Fragments ◽  
Dna Concentration ◽  
Large Molecular Weight ◽  
England Biolabs ◽  
Dna Precipitation ◽  
Storage Buffer

Protocol to precipitate extracted DNA from an aqueous solution to increase concentration or resolubilize in a different storage buffer. *Use isopropanol DNA precipitation if your DNA is suspended in a very large volume, if your DNA concentration is low, or you are trying to concentrate large molecular weight DNA fragments and remove smaller fragments. *Use ethanol DNA precipitation if you are trying to remove salt contamination or precipitate small DNA fragments. Additional Resources: New England Biolabs, "DNA Precipitation: Ethanol vs. Isopropanol". June 23, 2015 Green, Michael R. and Joseph Sambrook, "Precipitation of DNA with Isopropanol". doi:10.1101/pdb.prot093385Cold Spring Harb Protoc2017. Qiagen, "How can I precipitate genomic DNA using isopropanol?".

scholarly journals Comparison of DNA Concentration and Purity of Animal Blood Extracted using Different DNA Extraction Kits

2018 ◽  
Vol 2 ◽  
Author(s):  
Mohamed Sabri Esa ◽  
Nur Huda Faujan ◽  
Haitham Abdullah Rajab ◽  
Maryam Mohamed Rehan ◽  
Norlelawati Arifin ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Method ◽  
Bos Taurus ◽  
Dna Purification ◽  
Chain Reaction ◽  
Animal Blood ◽  
Dna Concentration ◽  
Dna Extraction Method ◽  
Polymerase Chain

The efficiency of DNA extraction from whole blood using appropriate method is very important for molecular analysis. Therefore, the aim of this study was to compare the purity and concentration of DNA extraction method from bovine (Bos taurus), chicken (Gallus gallus), and porcine (Sus scrofa) blood. The DNA of blood samples was extracted using three types of kit, namely InnuPREP Blood DNA Mini Kit, Wizard Genomic DNA Purification Kit, and QIAamp DNA Blood Mini Kit. The results showed that blood DNA extracted using QIAamp DNA Blood Mini Kit was found to be the most effective and consistently produced high concentrated and pure DNA for three animal samples. The purity of DNA ranged from 1.73 ± 0.05 Å to 1.94 ± 0.21 Å and the range of blood DNA concentration extracted using the QIAamp DNA were between 13.73 ± 2.11 and 25.01 ± 2.08 ng/?l. However, the blood DNA of porcine was not successfully extracted using InnuPREP Blood DNA Mini Kit and Wizard® Genomic DNA Purification Kit. These results were very crucial for the subsequent use of amplification using polymerase chain reaction (PCR) and to facilitate accurate detection in further analysis.

Indirect and direct damage to genomic DNA induced by 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin upon photodynamic action

2016 ◽  
Vol 20 (01n04) ◽  
pp. 331-336 ◽  
Author(s):  
Maria Bartolomeu ◽  
Šónia Coimbra ◽  
Ângela Cunha ◽  
Maria G.P.M.S. Neves ◽  
José A.S. Cavaleiro ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Genomic Dna ◽  
Antimicrobial Treatment ◽  
Photodynamic Inactivation ◽  
Bacterial Cells ◽  
Experimental Conditions ◽  
Dna Concentration ◽  
Transcription Process ◽  
Direct Exposure ◽  
Membrane Components

Photodynamic inactivation has been proposed as an efficient antimicrobial treatment for localized infections. Even though it is generally accepted that the cell wall and membrane components are the main targets of the photodynamic process, the importance of the nucleic acids as photodynamic targets is not yet fully understood. In this study, we investigated the photodamage of the genomic nucleic acids of the Gram negative bacterium Escherichia coli, using 5,10,15-tris(1-methylpyridinium-4-yl)-20-(pentafluorophenyl)porphyrin tri-iodide (Tri-Py[Formula: see text]-Me-PF) as photosensitizing agent. We tested, for the first time, the indirect photodamage effects on genomic DNA extracted from photosensitized bacteria and compared it with the direct effects on genomic DNA extracted from non-photosensitized cells, treated in otherwise similar experimental conditions. The results suggest that DNA does not seem to be a major target of photodynamic inactivation, once direct exposure to photosensitization does not damage DNA and does not significantly alter DNA concentration. The decrease in DNA concentration observed during the indirect exposure to photosensitization is directly related with the reduction of the concentration of bacterial cells. However, RNA synthesis was severely affected, once an indirect effect on proteins involved in the transcription process may cause a marked decrease in the RNA pool.

Paris I Dysfibrinogenemia: A Point Mutation in Intron 8 Results in Insertion of a 15 Amino Acid Sequence in the Fibrinogen γ-Chain

1993 ◽  
Vol 69 (03) ◽  
pp. 217-220 ◽  
Author(s):  
Jonathan B Rosenberg ◽  
Peter J Newman ◽  
Michael W Mosesson ◽  
Marie-Claude Guillin ◽  
David L Amrani
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Point Mutation ◽  
Genomic Dna ◽  
Comparative Sequence Analysis ◽  
Chain Gene ◽  
Molecular Defect ◽  
Nucleotide Position ◽  
Normal Individuals ◽  
Polymerase Chain

SummaryParis I dysfibrinogenemia results in the production of a fibrinogen molecule containing a functionally abnormal γ-chain. We determined the basis of the molecular defect using polymerase chain reaction (PCR) to amplify the γ-chain region of the Paris I subject’s genomic DNA. Comparative sequence analysis of cloned PCR segments of normal and Paris I genomic DNA revealed only an A→G point mutation occurring at nucleotide position 6588 within intron 8 of the Paris I γ-chain gene. We examined six normal individuals and found only normal sequence in this region, indicating that this change is not likely to represent a normal polymorphism. This nucleotide change leads to a 45 bp fragment being inserted between exons 8 and 9 in the mature γparis I chain mRNA, and encodes a 15 amino acid insert after γ350 [M-C-G-E-A-L-P-M-L-K-D-P-C-Y]. Alternative splicing of this region from intron 8 into the mature Paris I γ-chain mRNA also results after translation into a substitution of S for G at position γ351. Biochemical studies of 14C-iodoacetamide incorporation into disulfide-reduced Paris I and normal fibrinogen corroborated the molecular biologic predictions that two additional cysteine residues exist within the γpariS I chain. We conclude that the insertion of this amino acid sequence leads to a conformationallyaltered, and dysfunctional γ-chain in Paris I fibrinogen.

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