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Fastprep 24 automated homogenizer

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The FastPrep-24 is an automated homogenizer designed for rapid and efficient sample preparation. It utilizes a linear motion to agitate samples, enabling the disruption and homogenization of various biological materials, such as tissues, cells, and microorganisms.

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Lab products found in correlation

Ambion microbexpress bacterial mrna enrichment kit, by Thermo Fisher Scientific (3 mentions) Fastrna pro blue kit, by MP Biomedicals (3 mentions) Superscript first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Ribo zero magnetic kit, by Illumina (1 mentions) Agilent 2100 bioanalyzer, by Agilent Technologies (1 mentions) Synergy h4 microplate reader, by Agilent Technologies (1 mentions) Prism 6, by GraphPad (1 mentions)

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FastPrep-24 (832 citations) FastPrep (251 citations) FastPrep-24 homogenizer (191 citations) FastPrep homogenizer (75 citations) MP Fastprep-24 Automated Homogenizer (4 citations) FastPrep Automated Homogenizer (2 citations)

5 protocols using fastprep 24 automated homogenizer

1

Spore Formation Gene Expression Analysis

Cited 1 time
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RNAs from WT/EV, sleC mutant/EV, spo0A mutant/EV, spoVAC*/EV, spoVAC*/spoVAC, dpaAB mutant/EV, and dpaAB mutant/dpaAB strains grown for 18 h on 70:30 sporulation medium containing thiamphenicol (5 μg/ml) were extracted for quantitative real-time PCR (qRT-PCR) analyses of the spoVAC, spoVAD, spoVAE, dpaA, and dpaB transcripts. The RNA was extracted using a FastRNA Pro Blue kit (MP Biomedical) and a FastPrep-24 automated homogenizer (MP Biomedical). Contaminating genomic DNA was depleted using three successive DNase treatments, and mRNA enrichment was done using an Ambion MicrobExpress bacterial mRNA enrichment kit (Invitrogen), and samples were reverse transcribed as previously described (40 (link)).
Donnelly M.L., Fimlaid K.A, & Shen A. (2016). Characterization of Clostridium difficile Spores Lacking Either SpoVAC or Dipicolinic Acid Synthetase. Journal of Bacteriology, 198(11), 1694-1707.
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2

Transcriptional Regulation of Bacillus Sporulation

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RNA from WT, spo0A–, sigF–, spoIIQ–, sigE–, spoIIIA–, spoIIIAH–, and sigG−strains grown for 17 hrs on 70:30 sporulation media was extracted for qRT-PCR analyses of spoIIIAA transcript. RNA from WT, spo0A–, sigF–, spoIIQ–, sigE–, spoIIIA–, and sigG−strains grown for 25 hr on 70:30 sporulation media was extracted for qRT-PCR analyses of spoVT, CD1430, and spoVAD transcripts. RNA from WT/EV, spo0A–/EV, spoIIIAA–/EV, spoIIIA–/spoIIIA operon, and spoIIIA–/spoIIIAK167A operon complementation strains grown for 17 hr on 70:30 sporulation media was extracted for qRT-PCR analyses of spoIIIAA transcript. RNA was extracted using a FastRNA Pro Blue Kit (MP Biomedical) and a FastPrep-24 automated homogenizer (MP Biomedical). Contaminating genomic DNA was depleted using three successive DNase treatments and mRNA enrichment was done using an Ambion MICROBExpress Bacterial mRNA Enrichment Kit (Invitrogen). Samples were tested for genomic DNA contamination using quantitative PCR for rpoB. Enriched RNA was reverse transcribed using Super Script First Strand cDNA Synthesis Kit (Invitrogen) with random hexamer primers.
Fimlaid K.A., Jensen O., Donnelly M.L., Siegrist M.S, & Shen A. (2015). Regulation of Clostridium difficile Spore Formation by the SpoIIQ and SpoIIIA Proteins. PLoS Genetics, 11(10), e1005562.
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3

RNA Extraction and Sequencing of C. difficile

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RNA for RNA-Seq was extracted from WT, sigE−, sigK− and spoIIID−C. difficile after 18 hours of growth on 70:30 sporulation media as previously described (Fimlaid et al ., 2013 (link)). Briefly, RNA was extracted using a FastRNA Pro Blue Kit (MP Biomedical) and a FastPrep-24 automated homogenizer (MP Biomedical). Contaminating genomic DNA was depleted using three successive DNase treatments, and samples were tested for genomic DNA contamination using quantitative PCR for 16S rRNA and the sleC gene. DNase-treated RNA (5 µg) was mRNA enriched using a Ribo-Zero Magnetic Kit (Epicentre), and the quality of total RNA was validated using an Agilent 2100 Bioanalyzer.
RNA isolated for qRT-PCR was processed identically except that mRNA enrichment was done using an Ambion MICROBExpress Bacterial mRNA Enrichment Kit (Invitrogen). Reverse transcription of enriched RNA was done using the SuperScript® First Strand cDNA Synthesis Kit (Invitrogen) with random hexamer primers.
Pishdadian K., Fimlaid K.A, & Shen A. (2014). SpoIIID-mediated regulation of σK function during Clostridium difficile sporulation. Molecular microbiology, 95(2), 189-208.
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4

Tracking SIINFEKL Peptide Trafficking

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Six to eight week old mice were injected intradermally in each forearm with 30 μl (60 μl total) of 1 mM soluble SIINFEKL (10% TAMRA-labeled) or SLS-T (10% TAMRA-labeled). Mice were split into three end point groups: 4, 24, or 72 h (N = 4, 50% female, 50% male). For each group, mice were fluorescently imaged (IVIS Spectrum CT) under anesthesia, and then sacrificed at the indicated time point. Two additional mice were injected with 30 μl saline in each forearm (60 μl total) and sacrificed at 4 h to serve as controls. After sacrifice, axillary and brachial lymph nodes and spleens were harvested, imaged in IVIS Spectrum CT, and then placed into vials with 1.4 mm acid washed zirconium grinding beads (VWR). Lymph nodes were pooled into the same vial with 200 μl PBS, and spleens were placed in vial with 500 μl PBS. Organs were homogenized for 1 min in a FastPrep-24 Automated Homogenizer (MP Biomedicals) 0.150 μl of homogenate in PBS from each mouse's pooled lymph nodes or spleens were added to a 96-well-plate and analyzed for fluorescence using 557/583 nm excitation and emission reading on a plate reader (BioTek Synergy H4 Microplate Reader).
Tsoras A.N., Wong K.M., Paravastu A.K, & Champion J.A. (2020). Rational Design of Antigen Incorporation Into Subunit Vaccine Biomaterials Can Enhance Antigen-Specific Immune Responses. Frontiers in Immunology, 11, 1547.
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5

Tissue Distribution Quantification of Fluorescent Tracer

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At prescribed times post injection (p.i.), the tumor-draining and non-tumor-draining axillary and brachial lymph nodes, tumor or skin at the injection site, along with the spleen, lungs, liver, and kidneys were harvested and homogenized in D-PBS using 1.4 mm acid washed zirconium grinding beads (OPS Diagnostics LLC, Lebanon, NJ) with a FastPrep-24 Automated Homogenizer (MP Biomedicals, Santa Ana, CA). Tissue homogenate fluorescence was measured using a Synergy H4 BioTek plate reader and fluorescent tracer standard curves were made in individual tissue homogenates. Prism 6 (GraphPad, LaJolla, CA) was used to calculate exposure (as area under the curve from 1-72 hr post injection, AUC) from either the measured % injected or concentration of tracer in each tissue as well as the fluorescent tracer half-life of residence within the site of injection using one-phase decay least-squares regression. Error propagation analysis was used to calculate the standard error of AUC measurements. Concentrations are presented as the percent of tracer injection amount per tissue volume calculated from the respective tissue's weight and assuming a density of water.
Rohner N.A, & Thomas S.N. (2015). Melanoma growth effects on molecular clearance from tumors and biodistribution into systemic tissues versus draining lymph nodes. Journal of controlled release : official journal of the Controlled Release Society, 223, 99-108.
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