GSM7885941: IR48,rep2; Mus musculus; RNA-Seq
Mouse intestine was harvested and epithelial cells were eluted with 20 mM EDTA + 1 mM DTT and filtered through 100 and 40 um cell strainers. Cells were flash frozen and RNA was isolated using  miRNeasy Micro kit (Qiagen). Organoids were harvested using EDTA to remove matrigel and TrypLE to digest to single cell, at which point RNA was isolated using Zymo Direct-zol RNA Miniprep kit. Libraries were prepared using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina. Post amplification libraries were purified using Agencourt AMPure XP beads from Beckman Coulter. Libraries were validated using Agilent High Sensitivity DNA kit.