title: GSM5111175: KO-AhRf/f X Lgr5-EGFP-CreERT2 X Tomatof/f-H5; Mus musculus; RNA-Seq
library_construction_protocol: Subsequently, following transfer to chilled PBS-/-, crypts were mechanically separated from the connective tissue by vigorous vortexing. Cell suspensions were then filtered through a 40-µm mesh and Tomato-expressing cells (includes GFP+/Tom+ as well as GFP negative/Tom+) were collected using a MoFlo Astrios Cell Sorter (Beckman Coulter) or a Bio-Rad S3e Cell Sorter. Tomato positive cells represent colonic stem cells and their progeny. Single cell GEMs (Gel bead in EMulsion) and sequencing libraries were prepared using the 10x Genomics Chromium Controller in conjunction with the single-cell 3′ v3 kit. The cDNA synthesis, barcoding, and library preparation were then carried out according to the manufacturer's instructions.