scholarly journals Evaluation of a semi-automated, magnetic bead-based DNA extraction method for genetic fingerprinting of forensic casework samples

2008 ◽  
Vol 1 (1) ◽  
pp. 35-36 ◽  
Author(s):  
Barbara Haak ◽  
Andrea Porsche ◽  
Kai Vollack ◽  
Peter Zimmermann ◽  
Werner Pflug
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Magnetic Bead ◽  
Genetic Fingerprinting ◽  
Forensic Casework ◽  
Dna Extraction Method

scholarly journals MagnaExtract, a novel magnetic bead-based extraction method for the molecular detection of antimicrobial resistance genes in fresh water

2021 ◽  
Author(s):  
Rachel L. Byrne ◽  
Derek Cocker ◽  
Ghaith Alyayyoussi ◽  
Madalitso Mphasa ◽  
Mary Charles ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Dna Extraction ◽  
Resistance Genes ◽  
Water Samples ◽  
Extraction Method ◽  
Bacterial Isolate ◽  
Magnetic Bead ◽  
Dna Yield ◽  
Dna Extraction Method ◽  
Antimicrobial Resistance Genes

ABSTRACTBackgroundEnvironmental water samples are increasingly recognised as an important reservoir of antimicrobial resistance (AMR) genes. Polymerase chain reaction (PCR) and next generation sequencing (NGS) offer a potentially inclusive surveillance platform for a wide range of AMR genes. However, molecular methods are dependent upon the extraction of DNA of high yield and quality. Current options for DNA extraction from complex environmental matrices for downstream molecular applications are either expensive or low yielding. We present here a novel magnetic bead-based DNA extraction method, for the detection of antimicrobial resistance genes (ARGs) from river water in Malawi, named MagnaExtract.MethodsMagnaExtract involves initial filtration of 250ml freshwater, followed by an overnight incubation of the filter in 15ml buffered peptone water (BPW), common procedure in microbiology laboratories. 200µl is then taken for a boil (95°C) and spin step and mixed with magnetic beads to bind DNA. Following washes with ethanol, the DNA is eluted in nuclease-free water. To determine the effectiveness of this method, 98 freshwater samples were collected from two rivers in Southern Malawi, and DNA was isolated using the MagnaExtract method, two commercial Qiagen (Germany) kits; PowerWater and DNeasy Blood and tissue, alongside a boil and spin of BPW, and a boil and spin from bacterial isolate grown on agar media. All samples were screened with a high-resolution melt (HRM) PCR panel previously validated for the detection of third generation cephalosporin and carbapenem ARGs. We compared the DNA yield obtained using all extraction methods, as well as the identification of each ARG.ResultsDNA yield using MagnaExtract was statistically greater than both boil and spin methods and DNeasy Blood & Tissue (Qiagen, Germany). DNA yield was slightly lower than using PowerWater (Qiagen) but the difference was not statistically significant. MagnaExtract was the only method to identify ARGs in all 98 water samples compared with PowerWater (n=82), DNeasy (n=95) boilate of BPW (n=75) and boilate of bacterial isolate (n=87). The most commonly detected ARG was OXA-48 (n=93). In addition, we found overnight incubation in non-selective enrichment broth (BPW) to promote the growth of bacteria harbouring extended spectrum beta lactamase (ESBL) genes and reduction in the detection of carbapenemase genes.ConclusionThe MagnaExtract approach offers a simple, affordable, high yielding DNA extraction method for the detection of ARGs isolated from river water samples.

Sensitivity detection of Abacavir in human through SNP detection of HLA-B*5701 allele

2016 ◽  
Vol 5 (08) ◽  
pp. 4754
Author(s):  
Tanushree Mitra* ◽  
Shivshankar Kumdale ◽  
Sameer Chowdhary ◽  
Amol D. Raut
Keyword(s):  
Blood Sample ◽  
Dna Extraction ◽  
Genomic Dna ◽  
Extraction Method ◽  
Snp Detection ◽  
Dna Extraction Method ◽  
Sensitivity Detection

The main objective of this study was to make sure whether randomly taken 12 samples were sensitive to abacavir. The genomic DNA from 12 blood sample were extracted by phenol chloroform DNA extraction method, extracted genomic DNA were amplified and sequenced, thereafter SNPs were detected. Every sample had shown the presence of normal base at SNP position. This study indicated, those randomly taken 12 patients were sensitive to abacavir, so they can consume abacavir if they get infected with HIV.

Development of a rapid DNA extraction method and one-step nested PCR for the detection of Naegleria fowleri from the environment

2011 ◽  
Vol 45 (16) ◽  
pp. 5211-5217 ◽  
Author(s):  
Arine Fadzlun Ahmad ◽  
James Lonnen ◽  
Peter W. Andrew ◽  
Simon Kilvington
Keyword(s):  
Dna Extraction ◽  
Nested Pcr ◽  
Extraction Method ◽  
Naegleria Fowleri ◽  
Dna Extraction Method ◽  
One Step ◽  
Nægleria Fowleri

scholarly journals Choice of bacterial DNA extraction method from fecal material influences community structure as evaluated by metagenomic analysis

Microbiome ◽  
2014 ◽  
Vol 2 (1) ◽  
pp. 19 ◽  
Author(s):  
Agata Wesolowska-Andersen ◽  
Martin Bahl ◽  
Vera Carvalho ◽  
Karsten Kristiansen ◽  
Thomas Sicheritz-Pontén ◽  
...  
Keyword(s):  
Community Structure ◽  
Dna Extraction ◽  
Extraction Method ◽  
Metagenomic Analysis ◽  
Fecal Material ◽  
Bacterial Dna ◽  
Dna Extraction Method ◽  
Bacterial Dna Extraction

scholarly journals Evaluating the Impact of DNA Extraction Method on the Representation of Human Oral Bacterial and Fungal Communities

PLoS ONE ◽  
2017 ◽  
Vol 12 (1) ◽  
pp. e0169877 ◽  
Author(s):  
Anna Vesty ◽  
Kristi Biswas ◽  
Michael W. Taylor ◽  
Kim Gear ◽  
Richard G. Douglas
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Fungal Communities ◽  
Dna Extraction Method ◽  
The Impact

scholarly journals Optimized Protocol for Cyanobacterial 16S rRNA Analysis in Danube Delta Lakes

2021 ◽  
Author(s):  
Maria Iasmina Moza ◽  
Carmen Postolache
Keyword(s):  
16S Rrna ◽  
Dna Extraction ◽  
Phytoplankton Biomass ◽  
Extraction Method ◽  
Seasonal Difference ◽  
Danube Delta ◽  
16S Rrna Analysis ◽  
Dna Extraction Method ◽  
Optimized Protocol ◽  
Cyanobacterial Biomass

AbstractMolecular biology protocols have been more and more accessible to researchers for ecological investigations, however, these protocols always require optimization steps for the analysis of specific types of samples. The purpose of this study was to optimize a molecular protocol for the analysis of cyanobacterial 16S rRNA in Danube Delta shallows lakes. In this regard, several commercial DNA extraction kits were tested in comparison with potassium ethyl xanthogenate extraction method on different matrices. The obtained DNA was further used for 16S rRNA PCR optimization. Finally, an optimized protocol is proposed for the molecular analysis of cyanobacteria group in freshwater samples. The best DNA extraction method was the potassium xanthogenate extraction from dried cyanobacterial biomass. A dynamic in total genomic eDNA was observed, reflecting the seasonal difference in phytoplankton biomass from the studied lakes. The PCR protocol optimized by us can be successfully applied for the identification of a broad range of cyanobacterial genetic markers.

scholarly journals Improved DNA Extraction Method for Molecular Diagnosis from Smaller numbers of Cells

2014 ◽  
Vol 46 (3) ◽  
pp. 99-105 ◽  
Author(s):  
Seo Young Oh ◽  
Jeong Yeon Han ◽  
So Ra Lee ◽  
Hoon Taek Lee
Keyword(s):  
Dna Extraction ◽  
Molecular Diagnosis ◽  
Extraction Method ◽  
Dna Extraction Method
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