GSM3302172: SI IEC 4 weeks #1; Mus musculus; RNA-Seq
SI or colons were harvested from mice, flushed with PBS, cut open longitudinally, and incubated with 3 mM EDTA and 1 mM DTT for 30 minutes at 4C on a rocker. Supernatants were then passed through a 100 micron filter and centrifuged at 650xg for 5 minutes.  IECs were then resuspended in RNAzol, homogenized by passing repeatedly through a 25-gauge needle, and stored at -80C until RNA was isolated. RNA was isolated using a DirectZol RNA purification kit (Zymo Research). RNA was treated with Turbo DNAse (Thermo Fisher) for 30 minutes and purified using a RNA Clean and Concentrator kit (Zymo Research). 1 ug of total RNA was used for rRNA depletion with the RiboZero depletion kit to prepare RNA-sequencing library by using the TruSeq RNA sample prep kit (Illumina) according to manufacturer's instructions; 50-cycle single-end RNA sequencing was performed on a HiSeq 4000 (Illumina)