title: GSM7727986: WT_Organoid_Rep1; Mus musculus; RNA-Seq
library_construction_protocol: Epithelium was isolated from the duodenum of adult mice using a 5mM EDTA chelation for 30 minutes at 4°C followed by disaggregation with 4x TrypLE Select for 30 minutes at 37°C for the Lgr5_ApcKO samples while the others were incubated in Type 2 collagenase at 3mg/mL in HBSS for 30 minutes at 37°C. For the primary cells, ISCs were isolated using flow cytometry while the whole organoids were collected for RNA. The RNA was isolated using Trizol extraction followed by column purification using RNeasy Plus micro kit (Qiagen). Libraries were prepared using the NEBnext Ultra II non-directional library kit (New England Biolabs E7770S).