Qiashredder column

Manufactured by Qiagen

Sourced in Germany, United States, United Kingdom, Canada, Netherlands, Switzerland, France, Spain, Japan

The QIAshredder columns are a lab equipment product designed to homogenize and lyse cells or tissue samples. The columns use a unique shredding mechanism to disrupt the sample, improving the release of nucleic acids and other cellular components for subsequent processing and analysis.

Automatically generated - may contain errors

Lab products found in correlation

453 protocols using qiashredder column

mRNA Isolation and Quantitative RT-PCR from Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

For isolation of mRNA from Jurkat T-cells, the NucleoSpin^® RNA Kit (Macherey-Nagel, Düren, Germany) was used. To isolate mRNA from MT-2 cells, the RNeasy Mini Kit (Qiagen, Venlo, Netherlands) in combination with QIAshredder columns (Qiagen) was used. To isolate RNA from primary cells, the RNeasy Micro Kit (Qiagen) combined with QIAshredder columns was used. cDNA preparation from up to 5 µg of isolated RNA was performed with the SuperScript™ II Reverse Transcriptase (Thermo Fisher Scientific) combined with Random Hexamer Primers (Thermo Fisher Scientific) per manufacturer’s instructions. A technical triplicate of 200 ng of the obtained cDNA was subjected to quantitative real-time RT-PCR (qRT-PCR), employing the TaqMan^® Universal PCR Master Mix and a 7500 Real-Time PCR System (both Thermo Fisher Scientific). The primer/probe pairs and TaqMan Gene Expression Assays (Thermo Fisher Scientific) used to detect transcripts are listed in Table 1. Table 1 also lists the plasmids used to generate standard curves allowing for the quantification of transcripts. Data were evaluated with the 7500 Software v2.3 (Thermo Fisher Scientific). Every experiment was independently performed at least three times, and relative copy numbers (rcn) were calculated by normalization of respective transcript levels on those of β-actin.

Schnell A.P., Kohrt S., Aristodemou A., Taylor G.P., Bangham C.R, & Thoma-Kress A.K. (2022). HDAC inhibitors Panobinostat and Romidepsin enhance tax transcription in HTLV-1-infected cell lines and freshly isolated patients’ T-cells. Frontiers in Immunology, 13, 978800.

+ Open protocol

+ Expand

Transcriptional Analysis of Liver Samples

Check if the same lab product or an alternative is used in the 5 most similar protocols

For whole liver, the caudate lobe was snap frozen in liquid nitrogen and stored at−80 °C. RNA was extracted using QIAshredder columns and RNeasy mini columns (Qiagen) according to the manufacturer’s protocol, followed by quantification using the Nanodrop Spectrophotometer (Thermo Scientific); 0.5μg RNA was reverse-transcribed using SuperScript III (Invitrogen) according to the manufacturer’s protocol. Gene expression was calculated using the ΔΔCT method relative to housekeeping gene β-actin and Gapdh. For the in vitro phagocytosis assay, RNA was extracted using the QIAshredder columns and RNeasy mini columns (Qiagen), and 100 ng RNA were reverse transcribed using QuantiTect Reverse Transcription Kit (Qiagen) according to the manufacturer’s protocol. cDNA was then diluted to 1:10 with RNase Free water, prior to qPCR analysis. The following QuantiTect Primer Assays (Qiagen) were purchased: Tgf-β, Il10, Il10, Il6, Hmgb1, Sdf1/Cxcl12, Mcp1/Ccl2, Socs3, col1a2 and col3a1, Mmp2, Mmp7, a-SMA. Genes were analysed using the Quantifast SYBR Green PCR Kit (Qiagen) on an ABI 7500 Fast Real-Time System or a Roche LightCycler480 according to the manufacturer’s instructions.

Campana L., Starkey Lewis P.J., Pellicoro A., Aucott R.L., Man J., O'Duibhir E., Mok S.E., Ferreira-Gonzalez S., Livingstone E., Greenhalgh S.N., Hull K.L., Kendall T.J., Vernimmen D., Henderson N.C., Boulter L., Gregory C.D., Feng Y., Anderton S.M., Forbes S.J, & Iredale J.P. (2017). The STAT3-IL10-IL6 pathway is a novel regulator of macrophage efferocytosis and phenotypic conversion in sterile liver injury. Journal of immunology (Baltimore, Md. : 1950), 200(3), 1169-1187.

+ Open protocol

+ Expand

Quantifying Lipid Peroxidation in Flies

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peroxidized lipids were quantified using the lipid peroxidation kit (K739, BioVision) following the manufacturer’s instructions. Three pools of 20–40 young (2–4 days) and old (10–12 days) flies were lysed in 300 μL malondialdehyde (MDA) lysis buffer and homogenised on a QIAshredder column (QIAGEN) at 13,000 g for 10 min. Homogenates were diluted 1:4 before measurement. Total protein concentration was quantified using the Pierce BCA protein assay kit (Thermo Scientific) on 25 μL of undiluted lysate, following the manufacturer’s instructions. Colorimetric measurements were performed at 532 nm using a Biotek Synergy 2 plate reader. All measurements were performed in duplicate, and lipid peroxidation levels were normalized against total protein content of the sample.

Merkling S.H., Riahi H., Overheul G.J., Schenck A, & van Rij R.P. (2019). Peroxisome-associated Sgroppino links fat metabolism with survival after RNA virus infection in Drosophila. Scientific Reports, 9, 2065.

+ Open protocol

+ Expand

Quantifying IL-20 Signaling Pathway

Check if the same lab product or an alternative is used in the 5 most similar protocols

RNA was isolated from the treated Min6 β-cells and mouse islets or untreated db/db mouse islets using the RNeasy kit with the QiaShredder column (Qiagen, Denmark) following the manufacturer’s protocol. RNA was reverse transcribed to cDNA using cDNA Archive kit (Life Technologies, Denmark) according to the manufacturer’s protocol. qRT-PCR was performed on the Applied Biosystems Prism 7900HT real-time PCR machine and analyzed using SDS 2.4 software (Life Technologies, Denmark). The following primer/probes were purchased from Life Technologies: IL20 (Mm00445341_m1), IL20Ra (Mm00555504_m1), IL20Rb (Mm01232398-m1), IL22R (Mm00663697_m1), SOCS3 (Mm00545913_s1) and Rn18S (Hs99999901_s1). Relative transcript quantities were calculated by the standard curve method and normalized to the reference gene Rn18S.

Mayer C., Bergholdt R., Cucak H., Rolin B.C., Sams A, & Rosendahl A. (2015). Neutralizing Anti-IL20 Antibody Treatment Significantly Modulates Low Grade Inflammation without Affecting HbA1c in Type 2 Diabetic db/db Mice. PLoS ONE, 10(7), e0131306.

+ Open protocol

+ Expand

Subcellular Fractionation of NIH3T3 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

NIH3T3 cells (2 × 10⁸ cells/mL, 2 × 10 cm plates) were trypsinized and washed twice with ice cold PBS and lysed on ice for 15 min in a 100 µL cytoplasmic lysis buffer (10 mM HEPES pH 7.4, 10 mM KCl, 0.01 mM EDTA, 0.1 mM EGTA, 2 mM DTT, 5 mM Na₂VO₄, 0.1% NP40 and cOmplete, EDTA-free protease inhibitor cocktail (Sigma-Aldrich, Rehovot, Israel). Nuclei were pelleted using a benchtop microfuge (6000 rpm for 2 min) and the supernatant containing the cytoplasmic fraction was removed and treated with 2 M urea and 2% SDS and then boiled for 5 min. The nuclei were then washed twice in a 1 mL cytoplasmic lysis buffer to remove any cytoplasmic contaminant, and re-sedimented. Subsequently, the nuclei were lysed in 100 μL denaturing lysis buffer (2% SDS, 2 M urea, 8% sucrose, 1 mM NaF, and 5 mM Na₂VO₄). Genomic DNA was sheared by passage through a Qiashredder column (Qiagen, Crawley, UK) in a benchtop microfuge (6000 rpm for 2 min) and denatured by boiling for 5 min.

Katsman M., Azriel A., Horev G., Reizel Y, & Levi B.Z. (2022). N-VEGF, the Autoregulatory Arm of VEGF-A. Cells, 11(8), 1289.

+ Open protocol

+ Expand

Isolation of High-Quality RNA from J-Lat and CD4+ T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For RNA isolated from J-Lat 10.6 cells, cells first were passaged and split equally three times prior to isolation. Either J-Lat 10.6 or wild type for CCNT1 or knocked out for CCNT1 were each treated with TNFα (Peprotech, 300-01A) at 10 ng/mL or unstimulated in triplicate. For primary cell experiments, knockouts were performed similar to the “Primary CD4+ T cell activation Test,” and RNA was isolated after LRA treatment. In both J-Lat and primary CD4+ T cell isolation experiments, 0.1–2 × 10⁶ cells were isolated and resuspended in 350 μL of RLT Plus (Qiagen, 1053393) + 1% 2-mercaptoethanol (Millipore Sigma, M3148). Cells were frozen in buffer RLT Plus until RNA isolation. Thawed RLT lysates were then run over a QIAshredder column (Qiagen, 79654) and subsequently a gDNA eliminator column. A Qiagen RNeasy Plus Mini Kit was then used in order to obtain purified total RNA. RNA was submitted for TapeStation RNA assay or HighSense RNA assay (Fred Hutch Core Facilities Seattle, Washington, DC, USA), and RINe scores were all found to be ≥9.6.

Hafer T.L., Felton A., Delgado Y., Srinivasan H, & Emerman M. (2023). A CRISPR Screen of HIV Dependency Factors Reveals That CCNT1 Is Non-Essential in T Cells but Required for HIV-1 Reactivation from Latency. Viruses, 15(9), 1863.

+ Open protocol

+ Expand

RNA Isolation and cDNA Synthesis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were disrupted with lysis buffer containing 1% 2-mercaptoethanol and shredded with a QIAshredder column (Qiagen) [Li et al., 2011 (link)]. Total RNA from lysed cells was extracted and purified using an RNeasy kit (Qiagen). The concentration and purity of RNA was assessed using a NanoDrop (ThermoScientific). Synthesis of cDNA was performed using an iScript cDNA synthesis kit (Bio-Rad). The cDNA products were diluted 20-fold with DEPC treated water before use in the Taq-man assays. Primer/probe sets were premixed (Applied Biosystems).

Mooney S.M., Parsana P., Hernandez J.R., Liu X., Verdone J.E., Torga G., Harberg C.A, & Pienta K.J. (2015). The Presence of Androgen Receptor Elements Regulates ZEB1 Expression in the Absence of Androgen Receptor. Journal of cellular biochemistry, 116(1), 115-123.

+ Open protocol

+ Expand

Total RNA Extraction and cDNA Synthesis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total RNA extraction was performed using an RNeasy Plus Mini Kit (Qiagen) according to the manufacturer’s instructions after homogenization of each sample with a QIAshredder column (Qiagen). A DNase step was additionally performed during RNA isolation as described by the manufacturer. After RNA denaturation during 5 min at 65°C, 1 to 2 μg of total RNA was converted to cDNA using SuperScript™ III (Thermo Fisher Scientific) for 1 h at 50°C and the enzyme was then inactivated for 5 min at 85°C.

Girardot M., Bayet E., Maurin J., Fort P., Roux P, & Raynaud P. (2018). SOX9 has distinct regulatory roles in alternative splicing and transcription. Nucleic Acids Research, 46(17), 9106-9118.

+ Open protocol

+ Expand

Explant Culture of Human ACP Tumours

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Explant cultures were performed as previously described (Apps et al. 2018 (link)). Small pieces from three different human ACP tumours (approximately 1 mm³; four replicates per tumour) were placed on 0.2 μM Whatman filters (SLS) in 24-well plates containing 500 μL of media (DMEM-F12 (Gibco), 1% Pen/Strep (Sigma) and 1% FBS (PAA)) supplemented with either Vismodegib 100 μM (Selleckchem) or vehicle (DMSO) and medium was changed every 24 h. After 72 h, tumour pieces were passed through a Qiashredder column (Qiagen) and processed for total RNA extraction using the RNeasy Micro kit (Qiagen). Approximately 1 μg of total RNA was reverse-transcribed to cDNA using the Transcriptor First-Strand cDNA Synthesis Kit and random hexamers (Roche).

Carreno G., Boult J.K., Apps J., Gonzalez-Meljem J.M., Haston S., Guiho R., Stache C., Danielson L.S., Koers A., Smith L.M., Virasami A., Panousopoulos L., Buchfelder M., Jacques T.S., Chesler L., Robinson S.P, & Martinez-Barbera J.P. (2019). SHH pathway inhibition is protumourigenic in adamantinomatous craniopharyngioma. Endocrine-Related Cancer, 26(3), 355-366.

+ Open protocol

+ Expand

Efficient RNA Extraction from Transfected Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Three days post-transfection, Expi293-F or ExpiCHO-S cells were harvested by centrifugation and cell pellets were processed through a QIAshredder column (QIAGEN, Hilden, Germany). Total RNA extraction was carried out using RNeasy Mini Kit (QIAGEN) following the manufacturer’s instructions. RNA concentration was determined spectroscopically using a NanoDrop One (Thermo Fisher), ensuring pure RNA was isolated with a A260/280 ratio of 2.0.

Carrara S.C., Fiebig D., Bogen J.P., Grzeschik J., Hock B, & Kolmar H. (2021). Recombinant Antibody Production Using a Dual-Promoter Single Plasmid System. Antibodies, 10(2), 18.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!