GSM1644858: WT_intestinal stem cells; Mus musculus; RNA-Seq
Total RNA was extracted from 200K sorted GFPhi ISCs or GFPlow progenitors by pooling two to five 71-week old male mice per HFD and HFD-control using Tri Reagent (Life Technologies) according to the manufacturer's instructions, except for a overnight isopropanol precipitation at –20°C. From the total RNA, poly(A)+ RNA was selected using Oligo(dT)25-Dynabeads (Life technologies) according to the manufacturer’s protocol. Strand-specific RNA-seq libraries were prepared using the dUTP-based, Illumina-compatible NEXTflex Directional RNA-Seq Kit (Bioo Scientific) according to the manufacturer’s directions.