Improved Method for DNA Extraction and Purification from .

Methods Protoc

Laboratory of Biochemistry and Neurosciences, Department of Biology, University Hassan I, BP 577, 26000 Settat, Morocco.

Published: May 2019

(protozoa) is intensely investigated as a model organism, offering numerous advantages in comprehensive and multidisciplinary studies using morphologic or molecular methods. Since DNA extraction is a vital step of any molecular experiment, here a new mixed surfactant (Sodium dodecyl sulfate (SDS) 20%/Triton X-100) was adopted for effective DNA extraction from under an easy, fast protocol. The efficiency of this technique was then compared with three widely-used alternative techniques, namely the Chelex 100 matrix, Ammonium pyrrolidine dithiocarbamate (APD) complex and SDS-chloroform methods. DNA extraction was analyzed by pulsed-field gel electrophoresis, spectral measurement, fluorometry (Qubit), restriction enzyme digestion, and polymerase chain reaction. Data analysis revealed that the quantity and quality of the recovered DNA varied depending on the applied DNA extraction method. The new method (SDS 20%/Triton X-100) was the most efficient for extracting DNA from with high integrity and purity, affordable cost, less time, and suitability for molecular applications.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6632170PMC
http://dx.doi.org/10.3390/mps2020040DOI Listing

Publication Analysis

Top Keywords

dna extraction
20
methods dna
8
sds 20%/triton
8
20%/triton x-100
8
dna
7
extraction
5
improved method
4
method dna
4
extraction purification
4
purification protozoa
4

Similar Publications

Impact of automated nucleic acid extraction platforms on plasma Cytomegalovirus DNA loads quantitated by real-time PCR normalized to the 1st WHO international standard.

Enferm Infecc Microbiol Clin (Engl Ed)

January 2025

Microbiology Service, Clinic University Hospital, INCLIVA Health Research Institute, Valencia, Spain; CIBER de Enfermedades Infecciosas, Instituto de Salud Carlos III, Madrid, Spain; Department of Microbiology, School of Medicine, University of Valencia, Valencia, Spain. Electronic address:

Introduction: The extent to which commercially available nucleic acid extraction platforms impact the magnitude of Cytomegalovirus (CMV) DNA loads measured in plasma specimens by 1st WHO standard-normalized real-time PCR assays is uncertain.

Methods: This retrospective study compares the performance of Abbott m2000sp, Qiagen QIAsymphony SP, and KingFisher Flex platforms using plasma samples from allogeneic hematopoietic stem cell transplant recipients and plasma spiked with the CMV AD169 strain. The Abbott RealTime CMV PCR assay was used for CMV DNA quantitation.

View Article and Find Full Text PDF

Tuberculosis (TB) is the second deadliest infectious disease worldwide. Current TB diagnostics utilize sputum samples, which are difficult to obtain, and sample processing is time-consuming and difficult. This study developed an integrated diagnostic platform for the rapid visual detection of Mycobacterium tuberculosis (Mtb) in breath samples at the point-of-care (POC), especially in resource-limited settings.

View Article and Find Full Text PDF

Apical periodontitis microbiome association with salivary and serum inflammatory burden.

Int Endod J

January 2025

Department of Endodontics, Centre of Oral Clinical and Translational Sciences, Faculty of Dentistry, Oral and Craniofacial Sciences, Guy's Dental Hospital, King's College London, London, UK.

Aims: Apical Periodontitis (AP) involves complex interactions between the root canal microbiome and the host immune response, with potential risk of local and systemic inflammatory burden, however there is no evidence available regarding correlation between microbiome and inflammatory marker levels. This study aims to identify the microbiome of saliva, intracanal and blood samples in AP subjects and investigate the correlation between intracanal and blood microbiomes with serum inflammatory biomarker levels, and salivary microbiomes with salivary inflammatory biomarker levels.

Methodology: Saliva, Intracanal and blood samples were collected from AP patients undergoing root canal retreatment.

View Article and Find Full Text PDF

Our previous studies revealed the existence of a Universal Receptive System that regulates interactions between cells and their environment. This system is composed of DNA- and RNA-based Teazeled receptors (TezRs) found on the surface of prokaryotic and eukaryotic cells, as well as integrases and recombinases. In the current study, we aimed to provide further insight into the regulatory role of TezR and its loss in Staphylococcus aureus gene transcription.

View Article and Find Full Text PDF

Background: Synthesis of organic@inorganic hNFs is achieved by the coordination of organic compounds containing amine, amide, and diol groups with bivalent metals. The use of bio-extracts containing these functional groups instead of expensive organic inputs such as DNA, enzymes, and protein creates advantages in terms of cost and applicability. In this study, the application potentials (antioxidant, antibacterial, anticancer, guaiacol, anionic, and cationic dye degradation) of hybrid (organic@inorganic) nanoflowers (hNFs) synthesized with Cu and snakeskin (SSS) were proposed.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!