GSM3463030: Klf5-KO Lgr5+ ISC rep 1; Mus musculus; RNA-Seq
Proximal 1/3 of small intestine were resected, and crypts were isolated by villi scraping and 5mM EDTA incubation. Cells were dissociated with 4x TrpLE in DMEM for 45 minutes, and Lgr5-EGFPhi cells were sorted by flow cytometry. Total RNA was isolated using RNeasy Mini Kit (Qiagen) in combination with on-column DNase treatment. Library was prepared using the KAPA Stranded RNA-Seq kit with RiboErase (Kapa Biosystems) according to the manufacturer's instructions.