GSM9177871: Liver tissue, CD163KO, Western diet, rep E; Mus musculus; RNA-Seq
RNA was extracted from snap-frozen liver tissue. The tissue was homogenized in TRI reagent (T9424, Sigma-Aldrich) using the FastPrep-24 5G bead beating grinder. 1-bromo-3-chloropropane was added and vortexed thoroghly and incubated for 15min at RT followed by centrifugation (10000g, 10min, 4oC). The aqueous phase containing the RNA was mixed with cold 96% ethanol and loaded on a EconoSpin column (1920-250, Epoch Life Science) by centrifugation (13000g, 30sec, RT). After three washes with RPE buffer, the column was spinned dry for 2 minutes (13000g, RT) followed by 5 minutes to allow evapuration of ethanol. RNA was eluted by DEPC-treated water and centrifugation (16000g, 1min, RT). Libraries were prepared using NEBNext Ultra RNA Library Prep Kit for Illumina (New England Biolabs) and RNA was paired-end sequenced on the NovaSeq 6000 Sequencing System (Illumina).