GSM8625688: RNAseq_B6_HFD_1; Mus musculus; RNA-Seq
Total RNA was extracted from mouse eWAT tissue using the miRNeasy Mini Kit and QIAzol Lysis Reagent from QIAGEN, according to the manufacturer's protocol. The RNA was treated with RNase-Free DNase (QIAGEN) to remove any DNA. The RNA quality was assessed with an Agilent Bioanalyzer and the RNA 6000 Nano kit, with all samples showing a RIN score of 7 or higher. Ribosomal RNA was first removed with the NEBNext rRNA Depletion Kit (New England BioLabs). The libraries were then prepared using the NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England BioLabs), according to the manufacturer's guidelines. The yield of the libraries was assessed using the Qubit DNA High Sensitivity assay (Invitrogen), and quality control was conducted with the Agilent Bioanalyzer using the DNA 1000 kit (Agilent). Finally, the indexed libraries were pooled and sequenced on the NextSeq 500 platform (Illumina) with 75 bp single-end reads.