GSM3318503: Non-diabetic given the vehicle replicate 5; Mus musculus; RNA-Seq
Cells were disrupted with TRIzol (Qiagen, Hilde, Germany). RNA was isolated from TRIzol homogenates using the Direct-zol Kit (Zymo Research, Irvine, USA). RNA was quantified on Qubit (Thermo Fisher, Waltham, USA) and its quality was evaluated on the MultiNA bioanalyzer (Shimadzu, Kyoto, Japan). Some 200 ng of total RNA underwent ribosomal RNA depletion using the NEBNext® rRNA Depletion Kit followed by library construction using NEBNext® Ultra II™ Directional RNA Library Prep Kit for Illumina® (both from NEB, USA). Libraries were quantified on the MultiNA bioanalyzer (Shimadzu) and pooled to equimolar ratios for sequencing.