Tissuelyser 2 homogenizer

Manufactured by Qiagen

Sourced in Germany

The TissueLyser II is a high-throughput homogenizer designed for efficient disruption and homogenization of a wide range of sample types, including plant and animal tissues, bacteria, and yeast. It uses bead-beating technology to effectively break down samples and prepare them for downstream processing.

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Lab products found in correlation

Pierce bca protein assay kit, by Thermo Fisher Scientific (8 mentions) Rneasy mini kit, by Qiagen (4 mentions) 0.1 mm glass beads, by Qiagen (3 mentions) Powerbead tube, by Qiagen (3 mentions) Qscript cdna supermix, by Quanta Biosciences (3 mentions) Gel doc uv transilluminator system, by Bio-Rad (2 mentions) Gelred, by Biotium (2 mentions) Allprep dna rna mini kit, by Qiagen (2 mentions) Magna pure 96, by Roche (2 mentions) Abi 7500 fast real time pcr system, by Thermo Fisher Scientific (2 mentions) Rneasy plus universal kit, by Qiagen (2 mentions) Iscript cdna synthesis kit, by Bio-Rad (2 mentions) Lightcycler 480 sybr green 1 master, by Roche (2 mentions) Anti tubulin, by Santa Cruz Biotechnology (2 mentions) Odyssey infrared scanner, by LI COR (2 mentions) Lightcycler 96, by Roche (2 mentions) Rnalater, by Qiagen (2 mentions) Mobio powersoil kit, by Qiagen (1 mentions) Rneasy column kit, by Qiagen (1 mentions) Iscript advanced cdna synthesis kit for rt qpcr, by Bio-Rad (1 mentions)

33 protocols using tissuelyser 2 homogenizer

Stool DNA Extraction for Microbiome

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At the end of each study phase, subjects were instructed to collect a 2–4 g stool sample into a sealed container after recording food intake. Specimens were delivered within the first two hours after deposition and were stored at −80 °C until processing. All samples were identified and labeled with a random numeric code.
The MoBio PowerSoil kit (QIAGEN, Venlo, The Netherlands) was used to extract the DNA from stool samples according to the manufacturer’s instructions, including a pre-step of high shaking, to improve lysis, using a TissueLyser II homogenizer (QIAGEN). DNA was quantified using a NanoDrop ND-1000 spectrophotometer (Nanodrop Technologies, Wilmington, DE, USA).

Haro C., Guzmán-López M.H., Marín-Sanz M., Sánchez-León S., Vaquero L., Pastor J., Comino I., Sousa C., Vivas S., Landa B.B, & Barro F. (2022). Consumption of Tritordeum Bread Reduces Immunogenic Gluten Intake without Altering the Gut Microbiota. Foods, 11(10), 1439.

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UV-Crosslinking Liver Proteome-RNA Interactome

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100mg liver powder, on dry ice, was cross linked with UV (254 nm, 400 mJ/cm²) three times. AG dynabeads were washed twice in lysis buffer (50 mM Tris−HCl (7.4), 100 mM NaCl, 1% Igepal CA-630, 0.1% SDS, 0.5% sodium deoxcholate) and incubated with 10 μg antibody rotating at 4°C for 1 h then washed once in high salt buffer (50 mM Tris–HCl (7.4), 1 M NaCl, 1 mM EDTA, 1% Igepal CA-630, 0.1% SDS, 0.5% sodium deoxcholate) and twice in lysis buffer. Liver powder was resuspended in 1 ml lysis buffer (+Proteinase inhibitors & RNase inhibitors) and lysed using a TissueLyser II Homogenizer (Qiagen). 16 mg of protein was incubated at 37°C for 3 min with 10μl 1/1000 RNaseI (in lysis buffer) and 2 μl Turbo DNase then 3 min on ice. Samples were span at 18 000g for 10 min and the supernatant mixed with antibody bound beads and incubated at 4°C rotating for 1 h. Beads were washed twice with high salt buffer. 1/10 of beads were kept to assess immunoprecipitation efficiency and the rest treated with 50 μg PK to release RNA. RNA was purified using RNeasy kit column (Qiagen) and quantified using a nanodrop.

Jobbins A.M., Haberman N., Artigas N., Amourda C., Paterson H.A., Yu S., Blackford S.J., Montoya A., Dore M., Wang Y.F., Sardini A., Cebola I., Zuber J., Rashid S.T., Lenhard B, & Vernia S. (2022). Dysregulated RNA polyadenylation contributes to metabolic impairment in non-alcoholic fatty liver disease. Nucleic Acids Research, 50(6), 3379-3393.

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Faecal Sample Preparation for Virus Detection

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A volume of 2 mL of the faecal samples were taken for variations involving ultracentrifugation, while for other variations only 500 µL was taken to maximise handling efficiency and protect virus nucleic acid during extraction. Homogenization was carried out using TissueLyser II homogenizer (Qiagen) at 25 Hz for 2 min.

Vibin J., Chamings A., Collier F., Klaassen M., Nelson T.M, & Alexandersen S. (2018). Metagenomics detection and characterisation of viruses in faecal samples from Australian wild birds. Scientific Reports, 8, 8686.

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Quantifying Viral Cytopathic Effects

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Lung organoids were weighed and placed in 1 mL of DMEM supplemented with 1% heat-inactivated FBS, 1× L-glutamine before being homogenized in a Tissuelyser II Homogenizer (Qiagen) at 30 Hz for 6 min. Tissue homogenates were clarified by centrifugation at 5,000 × g for 5 min and then serially diluted 10-fold in DMEM supplemented with 2% heat-inactivated FBS and 2× penicillin–streptomycin. Sample volumes of 50 µL were added to 96-well plates of 95% confluent Vero76 cells in triplicate and incubated at 37 °C with 5% CO₂ before scoring for the presence of cytopathic effects.

Schuhenn J., Meister T.L., Todt D., Bracht T., Schork K., Billaud J.N., Elsner C., Heinen N., Karakoese Z., Haid S., Kumar S., Brunotte L., Eisenacher M., Di Y., Lew J., Falzarano D., Chen J., Yuan Z., Pietschmann T., Wiegmann B., Uebner H., Taube C., Le-Trilling V.T., Trilling M., Krawczyk A., Ludwig S., Sitek B., Steinmann E., Dittmer U., Lavender K.J., Sutter K, & Pfaender S. (2022). Differential interferon-α subtype induced immune signatures are associated with suppression of SARS-CoV-2 infection. Proceedings of the National Academy of Sciences of the United States of America, 119(8), e2111600119.

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Detection of Brucella spp. in Tissue Samples

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Frozen tissue samples of cases 5, 6 and 7 were submitted to PCR for the detection of Brucella spp. by hemi-nested PCR targeting an outer membrane protein gene of B. abortus [54 (link)].
The reactions were loaded as previously reported using B. suis bv 2 strain Thomsen as positive control and no template control as negative control.
Specifically, molecular detection was attempted on CNS, spleen, lung, liver, pre-scapular, tracheobronchial, pulmonary lymph nodes, tongue and skin ulcers, laryngeal tonsil and CSF of case 5; on CNS of case 6; on CNS, spleen, liver, lung and testes of case 7. For DNA extraction, tissue samples (30–50 mg) were physically disrupted using a TissueLyser II homogenizer (Qiagen, Hilden, Germany) by high-speed shaking in plastic tubes with stainless steel beads (5 mm in diameter). Genomic DNA was then extracted from the disrupted tissues with an AllPrep DNA/RNA Mini kit (Qiagen) according to the manufacturer’s instructions.
The PCR products were analyzed by electrophoresis on 2% agarose gel containing GelRed (Biotium, Fremont, California, USA), compared with molecular weight markers and subsequently photographed on a Gel-Doc UV transilluminator system (Bio-Rad, Hercules, California, USA).

Garofolo G., Petrella A., Lucifora G., Di Francesco G., Di Guardo G., Pautasso A., Iulini B., Varello K., Giorda F., Goria M., Dondo A., Zoppi S., Di Francesco C.E., Giglio S., Ferringo F., Serrecchia L., Ferrantino M.A., Zilli K., Janowicz A., Tittarelli M., Mignone W., Casalone C, & Grattarola C. (2020). Occurrence of Brucella ceti in striped dolphins from Italian Seas. PLoS ONE, 15(10), e0240178.

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High-throughput Microbiome DNA Extraction

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Mucosal swabs and tissue biopsies were placed into sterile Powerbead tubes preloaded with 0.1 mm glass beads (Qiagen, Hilden, Germany, Germantown, MD, USA) plus external lysis buffer in vitro diagnostic (200 µL, Roche Applied Science, Indianapolis, IN, USA). Tissues were homogenized at 30 Hz for 5 min using a Tissuelyser II homogenizer (Qiagen). Sample lysates were deposited into individual wells of 96 deep-well processing plates. DNA was subsequently extracted in high-throughput fashion using a MagNA Pure 96 instrument running a DNA and viral small volume-in vitro diagnostic extraction kit according to the manufacturer’s protocol (Roche). After extraction, a portion of the DNA was evaluated by ion torrent next generation sequencing or using the esophageal microbiome array (EMB). The remaining material was archived at −20 °C.

Cass S., Hamilton C., Miller A., Jupiter D., Khanipov K., Booth A., Pyles R., Krill T., Reep G, & Okereke I. (2021). Novel Ex Vivo Model to Examine the Mechanism and Relationship of Esophageal Microbiota and Disease. Biomedicines, 9(2), 142.

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RNA Extraction and cDNA Synthesis

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Tumor tissue adjacent to that used for cytogenetic analysis and histologic examination had been frozen and stored at −80°C from eight tumors (cases 1–8). Total RNA was extracted using miRNeasy kit, TissueLyser II homogenizer and Qiacube according to the manufacturer's recommendations (Qiagen Nordic, Stockholm, Sweden). For cDNA synthesis, 400–500 ng of total RNA were reverse-transcribed in a 20 μl reaction volume using iScript Advanced cDNA Synthesis kit for RT-qPCR according to the manufacturer's instructions (Bio-Rad Laboratories, Oslo, Norway). cDNA equivalent to 10 ng/μl of total RNA was used as template in subsequent real-time PCR assays. The Human Universal Reference Total RNA was used as control (Clontech Laboratories, Takara Bio Group, Saint-Germainen-Laye, France). According to the company's information, it is a mixture of total RNAs from a collection of adult human tissues, chosen to represent a broad range of expressed genes. Both male and female donors are represented.

PANAGOPOULOS I., GORUNOVA L., BJERKEHAGEN B., LOBMAIER I, & HEIM S. (2015). The recurrent chromosomal translocation t(12;18) (q14~15;q12~21) causes the fusion gene HMGA2-SETBP1 and HMGA2 expression in lipoma and osteochondrolipoma. International Journal of Oncology, 47(3), 884-890.

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Quantifying Metabolite Levels in Drosophila

Cited 2 times

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Protocols for protein, lipid, and carbohydrate measurements were performed as previously described (Kwon et al., 2015 (link); Ding et al., 2021 (link)). Four female flies were used for each replicate and a minimum of three replicates were measured for each sample group. Flies were homogenized in 200 ul 1X PBS with 0.1% Triton-X and Zirconium 1 mm Oxide Beads (Next Advance Lab Products, ZROB10) using TissueLyser II homogenizer (QIAGEN). Homogenate was incubated at 70°C for 10 minutes and the supernatant was collected after centrifugation at 3,000 g for 5 min. 5 ul of supernatant was applied to Pierce^™ BCA Protein Assay Kit (Thermo Scientific, 23227) for detecting protein levels. TAG and free glycerol levels were quantified from 20 ul supernatant using Triglycerides Reagent (Thermo Fisher Scientific^™ - TR22421) and Free Glycerol Reagent (Sigma-Aldrich, F6428), respectively. Free glycerol was subtracted from TAG values. Glucose levels were measured from 10 ul supernatant using Infinity Glucose Hexokinase Reagent (Thermo Fisher Scientific^™ - TR15421) or D-Glucose assay kit (Megazyme, K-GLUC). Trehalose levels were measured as for glucose but incubated with 0.4 ul trehalase (Megazyme, E-TREH). The amount of glucose was subtracted from trehalose read values. TAG, free glycerol, glucose, and trehalose levels were normalized to corresponding protein levels of each sample.

Liu Y., Dantas E., Ferrer M., Liu Y., Comjean A., Davidson E.E., Hu Y., Goncalves M.D., Janowitz T, & Perrimon N. (2023). Tumor Cytokine-Induced Hepatic Gluconeogenesis Contributes to Cancer Cachexia: Insights from Full Body Single Nuclei Sequencing. bioRxiv.

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Spleen Tissue RNA Extraction and qPCR

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Spleen tissues was homogenized using a TissueLyser II homogenizer (Qiagen). Total RNA was extracted from the homogenized samples using the RNeasy Plus Universal Kit from Qiagen (Hilden, Germany) following the manufacturer’s instructions. Genomic DNA contamination was eliminated through digestion with RNase-free DNase I (Qiagen). Reverse transcription (RT) was conducted using the iScript cDNA Synthesis Kit from Bio-Rad (Hercules, CA). Quantitative PCR (qPCR) was performed with the iTaq Universal SYBR Green Supermix from Bio-Rad on an ABI 7500 Fast Real-Time PCR System from Applied Biosystems (Waltham, MA). Relative gene expression was calculated, with 18S rRNA serving as the housekeeping gene for normalization. Triplicate reactions were run for each sample. Primer sequences are available upon request.

Alajoleen R.M., Oakland D.N., Estaleen R., Shakeri A., Lu R., Appiah M., Sun S., Neumann J., Kawauchi S., Cecere T.E., McMillan R.P., Reilly C.M, & Luo X.M. (2024). Tlr5 deficiency exacerbates lupus-like disease in the MRL/lpr mouse model. Frontiers in Immunology, 15, 1359534.

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Liver Cholesterol and Triglyceride Assay

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After rats were anesthetized by intraperitoneal injection of 3% pentobarbital (0.2 mL/100 g body weight), livers were taken out quickly. Liver tissues were put into isopropanol. Homogenates were manufactured using a TissueLyser-II homogenizer (QIAGEN, Germany), centrifuged at 3000 ×g, 4°C for 10 min, and then clear supernatants were collected. Total cholesterol (TC) and triglyceride (TG) in the liver tissue were determined with automatic biochemical analyzer (Olympus, Japan).

Yang Q.H., Xu Y.J., Liu Y.Z., Liang Y.J., Feng G.F., Zhang Y.P., Xing H.J., Yan H.Z, & Li Y.Y. (2014). Effects of Chaihu-Shugan-San and Shen-Ling-Bai-Zhu-San on p38 MAPK Pathway in Kupffer Cells of Nonalcoholic Steatohepatitis. Evidence-based Complementary and Alternative Medicine : eCAM, 2014, 671013.

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