Rneasy mini kit

Manufactured by Omni International

The RNeasy mini kit is a laboratory tool designed for the purification of total RNA from various biological samples. It utilizes a silica-based membrane technology to efficiently capture and elute RNA molecules, providing a reliable and consistent method for RNA isolation.

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Lab products found in correlation

Rneasy mini kit, by Qiagen (2 mentions) Bead ruptor 24, by Omni International (1 mentions)

2 protocols using rneasy mini kit

mRNA Sequencing and Differential Gene Expression in Tumor Samples

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For in vitro samples, cells were cultured as indicated and RNA was extracted using RNeasy mini kit (Qiagen) following manufacturer’s protocol. For tumor samples, tumors were dissected 2 weeks after injection. A 20 mg tissue from each sample was homogenized using Bead Ruptor 24 (Omni International) and RNA was processed using RNeasy mini kit following manufacturer’s protocol. Purified total RNA was cleaned up and mRNA was sequenced by NextSeq High Output. For analysis, reads were mapped to the mouse transcriptome (mm10, Ensembl annotations) using STAR (v2.7). FeatureCounts (v1.6) was used to count exonic reads (with -O option) and DESeq2 v1.20 (R v3.5.1) was used to normalize and compare samples. PCA plots were generated using the plotPCA function (DESeq2) after variance stabilization and dispersion estimation. Log-fold changes across all genes were used as input to iPAGE with the following parameters: ebins=9, max_p=0.005, and using MSigDB h gene sets (i.e. the Hallmarks gene set collection).

Zhu X.G., Chudnovskiy A., Baudrier L., Prizer B., Liu Y., Ostendorf B.N., Yamaguchi N., Arab A., Tavora B., Timson R., Heissel S., de Stanchina E., Molina H., Victora G.D., Goodarzi H, & Birsoy K. (2020). Functional genomics in vivo reveal metabolic dependencies of pancreatic cancer cells. Cell metabolism, 33(1), 211-221.e6.

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Cell Lysis and RNA Extraction Protocol

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Cell pellets stored at -80°C were resuspended with 600 μL buffer RLT 1% (v/v) 2-mercaptoethanol from the QIAGEN RNeasy Mini Kit (Cat No.: 74106) and then added into 2 mL OMNI prefilled ceramic bead tubes (SKU: 19–632). Tubes were loaded onto an OMNI Bead Mill Homogenizer (SKU:19-040E) for 3 beating cycles. For each cycle, samples were agitated at 5 m/s for 1 minute at 4 ^oC and then cooled on ice for 3 minutes between each cycle. The resulting lysate was clarified by centrifugation at 11,000 xg and then used for total RNA extraction with QIAGEN RNeasy Mini Kit (Cat No.: 74106) with on-column DNase digestion according to manufacturer’s instructions. Extracted total RNA for each sample was checked for purity and quantified with Invitrogen Qubit 2.0 Fluorometer (Cat No.: Q32866) and Qubit RNA HS Assay kit (Cat No.: Q32855) according to manufacturer’s instructions.

Li S., Li Y., Rushing B.R., Harris S.E., McRitchie S.L., Jones J.C., Dominguez D., Sumner S.J, & Dohlman H.G. (2021). Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein Asc1/RACK1. PLoS Genetics, 17(7), e1009640.

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