GSM4276601: AbsPro BR#2; Mus musculus; RNA-Seq
Mouse colons (caecum to rectum) were removed, flushed, and linearized and then dissociated with EDTA and mechanical force. Populations of Epcam+ cells (See manuscript for details) were bulk sorted on a BD FACS Aria Fusion using a 100 µm nozzle (20 PSI) at a flow rate of 2.0 with a maximum threshold of 5000 events/sec. The sample chamber and collection tubes were kept at 4 °C. Following exclusion of debris and singlet/doublet discrimination, cells were gated as demonstrated in the paper supplement. Populations were sorted into TRIzol (Invitrogen # 15-596-018) for downstream RNA isolation and sequencing. RNA was extracted from TRIzol samples using a Direct-zol RNA Micro-Prep kit (Zymo #11-330M) and associated guidelines. Sorted samples of each cell type were pooled as needed at the start of RNA preparations to ensure a minimum of 2,500 cells per sample. RNA sample quality and concentration was evaluated using an Aligent Bioanalyzer on an RNA high sensitivity pico chip. RNA samples were then pooled as needed to allow 1 ng library preps with Clontech Low Input Pico Kit (Takara #634940)
GEO Accession
GSM4276601