DNA isolation was performed using the MagNA Pure LC DNA Isolation Kit—Large Volume (Roche Diagnostics, Mannheim, Germany) following the manufacturer’s instructions, for which 500-μL aliquots of EDTA-anticoagulated blood samples were prepared. Extracted DNA samples were eluted in 200 μL MagNA Pure LC DNA Isolation Kit-Large Volume Elution Buffer. The Mutation Analysis Core Facility (MAF) of Clinical Research Center, Karolinska University Hospital (Stockholm, Sweden) provided the genotyping of SNPs of interest (and quality control) applying the Mass Array platform with iPLEX Gold Chemistry [104 (link)]. Successful genotyping rate exceeded 98 percent.
Magna pure lc dna isolation kit large volume
Manufactured by Roche
Sourced in Switzerland
The MagNA Pure LC DNA Isolation Kit—Large Volume is a laboratory equipment product designed to isolate DNA from various sample types. It utilizes magnetic bead-based technology to capture and purify DNA. The kit is suitable for processing larger sample volumes compared to standard DNA isolation kits.
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Lab products found in correlation
Magna pure lc dna isolation kit large volume elution buffer, by Roche (2 mentions)
Magna pure lc system, by Roche (2 mentions)
Nanodrop, by Thermo Fisher Scientific (2 mentions)
Qubit dsdna hs assay kit, by Thermo Fisher Scientific (2 mentions)
Massarray platform, by Labcorp (1 mentions)
Iplex gold chemistry, by Labcorp (1 mentions)
Magna pure 96 dna and viral nucleic acid small volume kit, by Roche (1 mentions)
Allprep dna rna micro kit, by Qiagen (1 mentions)
Nuclease free water, by Thermo Fisher Scientific (1 mentions)
Lightcycler faststart dna master hybprobe mix, by Roche (1 mentions)
Simpleprobe probes, by TIB Molbiol (1 mentions)
Abi prism 3130 genetic analyzer, by Thermo Fisher Scientific (1 mentions)
Bigdye terminator v3.1 cycle sequencing kit, by Thermo Fisher Scientific (1 mentions)
Magna pure lc, by Roche (1 mentions)
13 protocols using magna pure lc dna isolation kit large volume
1
DNA Isolation and Genotyping Protocol
2
DNA Isolation from Blood Samples
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DNA isolation was performed from ethylenediaminetetraacetic acid-anticoagulated blood samples using the MagNA Pure LC DNA Isolation Kit—Large Volume (Roche Diagnostics, Mannheim, Germany) according to the manufacturer’s protocol. The extracted DNA samples were eluted in MagNA Pure LC DNA Isolation Kit-Large Volume Elution Buffer (Roche Diagnostics) and stored in − 30 °C until measurements.
3
Polish Population Genomic DNA Sampling
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Human genomic DNA samples were derived from anonymous Polish unrelated volunteers who declared to be healthy. Samples were randomly selected from the “normal Polish population” genetic collection at the Biobank Lab, Department of Molecular Biophysics, University of Lodz. Genetic material for this collection was sampled in 2011 – 2012 within the EU-funded TESTOPLEK project. All subjects gave their written informed consent to participate in the study. This study was approved by the relevant regional ethical committee (Research Bioethics Commission, University of Lodz - Decision no. 8/KBBN-UŁ/II/2014 and Statement of the Research Bioethics Commission, University of Lodz from 17th of June, 2010) and all procedures were performed in accordance with the Declaration of Helsinki.
Saliva was collected into Oragene OG-500 DNA collection/storage receptacles (DNA Genotek, Kanata, Canada) and genomic DNA was subsequently isolated by the MagNA Pure LC DNA Isolation Kit - Large Volume (Roche, Basel, Switzerland) with final concentration normalized to 200 pg/μl [27 (link)]. A total of 190 samples were enrolled in the scanning study and other 380 in genotyping.
Saliva was collected into Oragene OG-500 DNA collection/storage receptacles (DNA Genotek, Kanata, Canada) and genomic DNA was subsequently isolated by the MagNA Pure LC DNA Isolation Kit - Large Volume (Roche, Basel, Switzerland) with final concentration normalized to 200 pg/μl [27 (link)]. A total of 190 samples were enrolled in the scanning study and other 380 in genotyping.
4
DNA Isolation from Blood Samples
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Following the manufacturer’s instructions, we used a MagNA Pure LC system (Roche Diagnostics, Basel, Switzerland) with a MagNA Pure LC DNA Isolation Kit–Large Volume to isolate DNA from EDTA-anticoagulated blood samples.
5
DNA Isolation and Genotyping Methodology
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DNA isolation and genotyping method were exposed elsewhere [28 (link)]. In brief, DNA preparation was carried out from EDTA-anticoagulation blood samples on the day of sample collection. From plasma samples, DNA were isolated using MagNA Pure LC DNA Isolation Kit–Large Volume (Roche Diagnostics, Basel, Switzerland) according to the manufacturer’s instructions. Genotyping was performed on a MassARRAY platform (Sequenom Inc., San Diego, CA, USA) with iPLEX Gold chemistry by the Mutation Analysis Facility service provider (MAF, Karolinska Institute, Solna, Sweden). Validation, concordance analysis, and quality control were conducted by the Facility according to their protocols.
6
Genomic DNA Extraction and T. parva Detection
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Genomic DNA was extracted from in vitro cell cultures and blood material using MPLC or MP96 instruments with their respective MagNa Pure LC DNA Isolation Kit – Large Volume or MagNA Pure 96 DNA and Viral Nucleic Acid Small Volume Kit (Roche Diagnostics, Mannheim, Germany), according to manufacturer’s instructions. Eluted DNA was tested for T. parva using the Hybrid II assay (Pienaar et al., 2011 (link)). Molecular biology work was done in a South African National Accreditation System (SANAS) accredited (V0017) and Department of Agriculture, Land Reform and Rural development (DALRRD) (DAFF-30) approved laboratory.
7
DNA Extraction from Blood Samples
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A MagNA Pure LC system (Roche Diagnostics, Basel, Switzerland) with a MagNA Pure LC DNA Isolation Kit-Large Volume was used to isolate DNA from the blood sample according to the instructions of the manufacturer. Extracted DNA was eluted in a 200 μl MagNA Pure LC DNA Isolation Kit-Large Volume elution buffer.
8
DNA Extraction from Whole Blood
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DNA isolation was performed from ethylenediaminetetraacetic acid-anticoagulated blood samples using the MagNA Pure LC DNA Isolation Kit—Large Volume (Roche Diagnostics, Mannheim, Germany) according to the manufacturer’s protocol. The extracted DNA samples were eluted in a MagNA Pure LC DNA Isolation Kit-Large Volume Elution Buffer (Roche Diagnostics, Basel, Switzerland) and stored at −30 °C until measurements were carried out.
9
Genetic Analysis of Madurella Mycetomatis
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Individuals presenting at Mycetoma Research Center, Khartoum between 2001 and 2008, were eligible for inclusion, when the diagnosis of Madurella mycetomatis mycetoma was confirmed. People living in the same endemic regions were included as controls. The demographic features are given in Table 1 . Retrospectively, genotypes were determined of 112 Madurella mycetomatis infected patients and 103 healthy endemic controls, matched for sex and age. Whole blood was stored at -80°C until processing. DNA was isolated from whole blood of these subjects using the MagNA Pure LC DNA Isolation kit—Large Volume (Roche Diagnostics Nederland BV, Almere, the Netherlands) according to the manufacturer’s instructions. DNA was stored at -20°C until processing. Tissue of the foot and of the grain was obtained in 1998 from Sudanese subjects, infected with M. mycetomatis.
10
Nucleic Acid Extraction Protocols for Malaria
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Human blood samples (100 µl) from The Gambia (both years) were extracted using the Boom method (Boom et al., 1990 (link)) and eluted in 50 µl of nuclease-free water (Ambion, UK). One hundred microliters of whole blood samples from the Burkina Faso studies were processed using the MagNAPure Total Nucleic Acid Isolation Kit – High Performance (Roche, The Netherlands) and eluted in 50 µl of nuclease-free water. Asexual parasite samples were extracted using the MagNAPure LC DNA Isolation Kit – Large Volume (Roche) and eluted in 100 µl of nuclease-free water. Enriched gametocyte DNA was extracted by an AllPrep DNA/RNA Micro kit (Qiagen) and eluted in 100 µl of elution buffer according to the manufacturer’s guide.
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