Total RNA from distal gut was isolated by homogenization of ~100 mg of tissue in TRIzol® Reagent (Ambion by Life Technologies, Carlsbad, CA, USA), using 3 mm tungsten carbide beads and a TissueLyser II Disruption System (Qiagen GmbH, Hilden, Germany). Following isolation, the RNA was quantified by spectrophotometry (NanoDrop Technologies, Wilmington, DE, USA), had integrity confirmed by electrophoresis (Agilent Technologies, Santa Clara, CA, USA) and was then stored at − 80 °C for later processing.
Tissuelyser 2 disruption system
Manufactured by Qiagen
Sourced in Germany
The TissueLyser II Disruption System is a high-throughput sample preparation device designed for efficient disruption and homogenization of a wide range of sample types, including plant, animal, and microbial tissues. The system utilizes bead-beating technology to effectively break down samples, preparing them for downstream processing and analysis.
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Lab products found in correlation
5 protocols using tissuelyser 2 disruption system
1
Total RNA Extraction from Distal Gut
2
Inguinal Mammary Gland RNA Isolation
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Total RNA from the inguinal mammary gland was isolated by homogenization of ~ 100 mg of tissue in TRIzol® Reagent (Ambion by Life Technologies, Carlsbad, CA, USA), using 3 mm tungsten carbide beads and a TissueLyser II Disruption System (Qiagen GmbH, Hilden, Germany). Following isolation, the RNA was quantified by spectrophotometry (NanoDrop Technologies, Wilmington, DE, USA) and its integrity was confirmed by electrophoresis (Agilent Technologies, Santa Clara, CA, USA). All RNA samples had a RIN number > 7.8, meeting the criteria for RNA-seq.
3
Gill Tissue RNA Extraction Protocol
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Total RNA extraction was performed on individual gill samples (n = 64), including the gill arch and full-length filaments. The RNA was isolated by homogenization of ~100 mg of gill tissue in TRIzol® Reagent (Ambion by Life Technologies, Carlsbad, CA, United States), using 3 mm tungsten carbide beads and a TissueLyser II Disruption System (Qiagen GmbH, Hilden, Germany). Afterwards, the RNA was quantified by spectrophotometry (NanoDrop Technologies, Wilmington, DE, United States), with the RNA integrity assessed by electrophoresis (Agilent Technologies, Santa Clara, CA, United States). The individual RNA samples were subsequently pooled to generate 4 biological replicates per each group, with an equimolar contribution of RNA from 4 gill samples to each pool, yielding 16 RNA pools in total (4 groups x 4 RNA pools x 4 gill samples per pool). The 4 gill samples per pool originated from 2–4 fish.
4
Inflammatory Cytokine Quantification in Mouse Skin
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The fresh mice back skin was individually weighed and placed into separate tubes containing 1 mL of phosphate buffer saline (PBS) with protease inhibitor. The skin tissue was homogenized using the Tissuelyser II Disruption System (Qiagen, CA, USA) at a frequency of 30 per second for 10 minutes, followed by centrifugation at 12,000 RPM at 4 ◦C for 10 minutes. The levels of inflammatory cytokines including TNF-α, IL-23 and IL-6 in the supernatant were quantified using BioLegend ELISA kits (Biolegend, San Diego, CA, USA) according to the manufacturer’s instructions.
5
Comprehensive Gill Tissue RNA Extraction
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Total RNA extraction was performed on individual gill transverse sections (45 gills × 3 sections) after removing the arch tissue and leaving only full-length filaments for further processing (Figure 1 ). Briefly, the RNA was isolated by homogenization of ∼100 mg of gill tissue in TRIzol® Reagent (Ambion by Life Technologies, Carlsbad, CA, United States), using 3 mm tungsten carbide beads and a TissueLyser II Disruption System (Qiagen GmbH, Hilden, Germany). Following isolation, the RNA was quantified by spectrophotometry (NanoDrop Technologies, Wilmington, DE, United States) and its integrity was confirmed by electrophoresis (Agilent Technologies, Santa Clara, CA, United States). The three individual RNA samples that originated from the same gill were then pooled to generate a single RNA sample per gill tissue (n = 45 RNA pools in total), with an equimolar contribution of RNA from dorsal, medial and ventral regions of the gill to each pool. All but one pooled gill RNA samples had a 260/280 ratio > 1.8 and RIN number > 9.3, thus meeting the criteria for RNA-sequencing. The sample with degraded RNA was eliminated from further processing.
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