Total antioxidant capacity (ferric reducing antioxidant power, FRAP) was extracted in ice-cold 80% ethanol using a MagNALyser (Roche, Vilvoorde, Belgium) and measured using Trolox as a reference [55 (link)]. Plant materials were extracted in 1mL 80% ethanol using MagNALyser for ascorbate and glutathione measurement (Roche, Vilvoorde, Belgium). HPLC was used to measure reduced ascorbate (ASC) and reduced glutathione (GSH). Total ascorbate (ASC + DHA) and glutathione (GSH + GSSG) concentrations were evaluated after dithiothreitol (DTT) reduction, as previously reported [13 (link)]. Polyphenols and flavonoids were extracted by homogenizing fresh plant materials in 80% ethanol and centrifuged for 15 min at 5000 rpm. The clear extract was then used to determine the total phenolic and flavonoid content using the Folin-Ciocalteu and aluminum chloride tests.
Magna lyser
Manufactured by Roche
Sourced in Switzerland, Belgium, Germany, United States, United Kingdom, Austria, Italy, Spain, Netherlands, China
The MagNA Lyser is a high-speed bead-beating instrument designed for efficient disruption and homogenization of various sample types, including tissues, cells, and microorganisms. It utilizes a rapid shaking motion to facilitate mechanical lysis and extract nucleic acids, proteins, or other target molecules from the samples. The MagNA Lyser is a versatile tool for sample preparation in a wide range of life science applications.
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Lab products found in correlation
Rneasy mini kit, by Qiagen (27 mentions)
Trizol reagent, by Thermo Fisher Scientific (27 mentions)
Magna lyser green beads, by Roche (25 mentions)
Trizol, by Thermo Fisher Scientific (14 mentions)
Nanodrop, by Thermo Fisher Scientific (13 mentions)
Nanodrop spectrophotometer, by Thermo Fisher Scientific (11 mentions)
Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (9 mentions)
Spd m10avp, by Shimadzu (8 mentions)
Magmax viral rna isolation kit, by Thermo Fisher Scientific (7 mentions)
Qiaamp dna mini kit, by Qiagen (6 mentions)
Nanodrop nd 1000, by Thermo Fisher Scientific (6 mentions)
Rnase free dnase set, by Qiagen (6 mentions)
Synergy mx, by Agilent Technologies (6 mentions)
Nanodrop 1000, by Thermo Fisher Scientific (6 mentions)
Nanodrop 2000, by Thermo Fisher Scientific (6 mentions)
Nanodrop 1000 spectrophotometer, by Thermo Fisher Scientific (6 mentions)
Qubit fluorometer, by Thermo Fisher Scientific (6 mentions)
Magna pure lc, by Roche (5 mentions)
Rneasy kit, by Qiagen (5 mentions)
Magna lyser green beads tubes, by Roche (5 mentions)
388 protocols using magna lyser
1
Antioxidant Profiling of Plant Extracts
2
Quantifying Uterine Cytokines in HMA Mice
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The uterus from HMA mice at 18.5 dpc was collected and the pups were carefully removed. The uterine wall was flash frozen with liquid nitrogen. To ensure adequate representation of the uterine tissue in subsequent cytokine analysis, the uterine tissue was ground while frozen with mortar and pestle. Fifty milligram of tissue was then added to homogenization buffer in a Magnalyser tube and homogenized twice for 30 s at 6,500 rpm using a Magnalyser (Roche, Basel, Switzerland). Protein concentrations were quantified using the Pierce BCA assay kit and the protein concentrations were diluted and normalized for cytokine quantification using the V-PLEX Proinflammatory Panel 1 Mouse Kit (Meso Scale Discovery, Rockville, Maryland) following manufacturer's protocol with the help of the Emory Multiplexed Immunoassay Core (Emory University, Atlanta, GA).
3
Comprehensive Antioxidant Analysis in Plants
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TAC (ferric reducing antioxidant power, FRAP) was measured in ice-cold 80% ethanol with a MagNALyser (Roche, Vilvoorde, Belgium) and quantified using Trolox as a reference, as reported by Benzie and Strain (1999) (link). Plant samples were extracted in 80% ethanol and measured for ascorbate and glutathione using a MagNALyser (Roche, Vilvoorde, Belgium). Reduced ascorbate (ASC) and glutathione (GSH) were measured using HPLC. To extract phenols and flavonoids, fresh plant materials were homogenized in 80% ethanol before being centrifuged at 5000 rpm for 15 minutes. The clear extract was then used to measure the phenols and flavonoid concentrations using the Folin-Ciocalteu and aluminum chloride assays, respectively (AbdElgawad et al., 2023 (link)). Tocopherols were extracted with hexane (100 mg FW in 6 ml hexane) and centrifuged for 15 minutes at 14,000 g. The extracts were dried (Labconco, Kansas, USA) and resuspended in hexane. HPLC (Shimadzu, Hertogenbosch, the Netherlands) was used to separate and quantify tocopherols under normal phase conditions. Particil Pac 5 mm column material, length 250 mm, i.d. 4.6 mm, and internal standard dimethyl tocol (DMT) (5 ppm). The Shimadzu Class VP 6.14 software was used to examine the data.
4
Penaeus monodon Broodstock Mortality Investigation
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In November 2012, a batch of 414 wild Penaeus monodon broodstock were caught by net and trawl in Joseph Bonaparte Gulf in northern Australia and transported to a commercial hatchery in North Queens land. Upon arrival, the shrimp were bathed in a prophylactic solution containing formalin and povidine-iodine before being placed into concrete raceways. Mortalities began to occur from Day 1 poststocking, increasing progressively to 81.4% by Day 12.
Due to the high mortality rate, tissues for histology, bacteriology and PCR were sampled from 3 moribund broodstocks by the Queensland Department of Agriculture, Fisheries and Forestry (QDAFF). Cephalothoraxes were fixed in Davidson's AFA fixative, processed for histology, and tissue sections were stained with either haematoxylin & eosin or Brown & Brenn's Gram stain using standard procedures (Lightner 1996) . Preliminary analysis suggested the presence of a yellow-head-complex virus.
Three samples of gill and epidermis, fixed in ethanol, were submitted to the CSIRO AAHL Fish Diseases Laboratory, Geelong, Victoria, for confirmatory testing. Samples were homogenised in 600 µl AVL buffer in MagNA Lyser green bead tubes using the MagNA Lyser (Roche) and clarified by centrifugation at 10 000 × g for 5 min. RNA was extracted from 140 µl using the QIAmp Viral RNA Mini Kit (QIAGEN) and eluted in a final volume of 60 µl with AVE buffer.
Due to the high mortality rate, tissues for histology, bacteriology and PCR were sampled from 3 moribund broodstocks by the Queensland Department of Agriculture, Fisheries and Forestry (QDAFF). Cephalothoraxes were fixed in Davidson's AFA fixative, processed for histology, and tissue sections were stained with either haematoxylin & eosin or Brown & Brenn's Gram stain using standard procedures (Lightner 1996) . Preliminary analysis suggested the presence of a yellow-head-complex virus.
Three samples of gill and epidermis, fixed in ethanol, were submitted to the CSIRO AAHL Fish Diseases Laboratory, Geelong, Victoria, for confirmatory testing. Samples were homogenised in 600 µl AVL buffer in MagNA Lyser green bead tubes using the MagNA Lyser (Roche) and clarified by centrifugation at 10 000 × g for 5 min. RNA was extracted from 140 µl using the QIAmp Viral RNA Mini Kit (QIAGEN) and eluted in a final volume of 60 µl with AVE buffer.
5
Bacterial Profiling of Fecal Samples
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Stool samples were homogenized, using the Roche MagNA Lyser (Roche Inc, Basel, Switzerland). DNA was extracted from the homogenized fecal isolates, using the QiaAmp PowerFecal DNA kit (Hilden, Germany). Bacterial profiles were determined by broad-range amplification and sequence analysis of 16S rRNA genes, following our previously described methods [30 (link),31 (link)]. In brief, amplicons were generated, using primers that target approximately 400 base pairs of the V3V4 variable region of the 16S rRNA gene. PCR products were normalized, using a SequalPrepTM kit (Invitrogen, Carlsbad, CA, U.S.A.), pooled, lyophilized, purified and concentrated, using a DNA Clean and Concentrator Kit (Zymo, Irvine, CA, U.S.A.). Pooled amplicons were quantified, using Qubit Fluorometer 2.0 (Invitrogen, Carlsbad, CA, U.S.A.). The pool was diluted to 4nM and denatured with 0.2 N NaOH at room temperature. The denatured DNA was diluted to 15 pM and spiked with 25% of the Illumina PhiX control DNA prior to loading the sequencer. Illumina paired-end sequencing was performed on the MiSeq platform with versions v2.4 of the Miseq Control Software and of MiSeq Reporter, using a 600 cycle version 3 reagent kit.
6
Protein Extraction and Immunoblotting
7
Molecular Identification of Fungal Strains
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Candida cultures used for the molecular identification of the strains were grown on yeast-peptone D-glucose (YPD) for 2 days, and DNA was extracted using the Masterpure™ Yeast DNA Purification Kit (Epicentre Biotechnol., Madison, USA) per the manufacturer’s instructions [42 (link)]. Mould reference strains (Aspergillus only) and clinical isolates (Aspergillus, Fusarium, Lichtemia, Rhyzopus, Scedosporium) were cultivated on standard minimal nitrate medium [43 (link)]. DNA extractions were carried out at the University of Debrecen (Department of Biotechnology and Microbiology). gDNA was isolated from liquid cultures grown in minimal medium at 37 °C (A. fumigatus, A. niger), 25 °C (A. terreus, A. lentulus, A. flavus) and 30 °C (A. welwitschiae) at 220 rpm for 18 h. The mycelium was disrupted by using Roche MagNa Lyser (Roche Diagnostics, Risch-Rotkreuz, Switzerland) and gDNA was isolated using the Genomic DNA Purification Kit (Thermo Scientific, Maryland, USA) per the manufacturer’s instructions.
8
High-Throughput DNA Extraction from Whole Blood
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DNA purification steps were performed in a class II laminar air-flow cabinet to avoid environmental contamination. Spiked EDTA-WB samples were disrupted (2000 rpm, 1 min) by Roche MagNa Lyser (Roche Diagnostics, Risch-Rotkreuz, Switzerland), and DNA was extracted along with the ENCs using the High Pure Viral Nucleic Acid Large Volume Kit (Roche Applied Science) according to manufacturer’s instructions. To obtain technical duplicates every sample extractions were performed in parallel. The elution volumes were adjusted to 15 μl and pooled.
9
Cytokine Profiling in Infected Mice
10
Isolation and Identification of Mycobacterium bovis
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Tissue samples were kept at −30°C until processing. Tissues (between 10 and 25 mg) were macerated with 1.5 ml of sterile distilled water in a MagNA Lyser apparatus (Roche,
Penzberg, Germany), decontaminated using the Petroff method [19 (link)] and cultured in Stonebrink culture medium tubes at 37°C for up to 90 days. Colonies
suggestive of M. bovis were submitted to conventional PCR with the Mb.400 primers, which were used to amplify a fragment of 400 bp flanking the region of differentiation 4
(RD4) specific to M. bovis [21 (link)].
Penzberg, Germany), decontaminated using the Petroff method [19 (link)] and cultured in Stonebrink culture medium tubes at 37°C for up to 90 days. Colonies
suggestive of M. bovis were submitted to conventional PCR with the Mb.400 primers, which were used to amplify a fragment of 400 bp flanking the region of differentiation 4
(RD4) specific to M. bovis [21 (link)].
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