GSM8121004: Rxra cKO FBS HFSCs 1, RNA-seq; Mus musculus; RNA-Seq
To resuspend HFSCs, 3D cultures were washed with sterile PBS, dissociated into single cell suspensions using TrpLE-Express (Gibco) for 15min at 37°C. After filtering, total RNA was collected in Trizol and purified using the Direct-zol RNA MiniPrep kit (Zymo Research) according to the manufacturer's instructions. DNase treatment was performed to remove genomic DNA (RNase-Free DNase Set, Qiagen). Sample quality was determined using an Agilent 2100 Bioanalyzer; all samples for sequencing had RNA integrity (RIN) numbers >8. cDNA library construction was performed by the Rockefeller University (RU) Genomic Core facility.