GSM4564588: Mm_EPI_HFD_WT_002; Mus musculus; RNA-Seq
Total RNA was isolated from cells or tissues using TRIzol reagent (Thermo Fisher) followed by a Direct-zol RNA MiniPrep kit (Zymo Research) according to the manufacturer's instructions. 400ng of total RNA was used as input for RNA-seq Library construction, which consisted of rRNA removal via the NEBNext rRNA depletion kit (E6310X) and conversion to cDNA using the NEBNext Ultra II Non-Directional RNA Second Strand Synthesis Module (E6111L), both per manufacturer's instructions. cDNA was diluted 1:15 and subsequently tagmented and amplified for 12 cycles using the Nextera XT DNA Library Preparation Kit (Illumina FC-131). Individual libraries were QC'd by Qubit and Agilent Bioanalyzer, pooled, and sequenced at a final loading concentration of 1.8 pM on a NextSeq500 (36bp x 36bp paired-end reads).