title: GSM6460331: Colon_PdgfraLo_Rep1; Mus musculus; RNA-Seq
library_construction_protocol: Mice were sacrificed and intracardially perfused with PBS, tissue was harvested, washed and the outer muscle layer was carefully removed with tweezers under a stereomicroscope. Tissue pieces were incubated twice in 10mM EDTA in HBSS +10mM HEPES + 5%FBS for 30min under 300rpm shaking at 37°C, washed and digested with 3mg/ml collagenase2 for 20min at 37°C. Filtered cell suspension was  washed  before staining procedures and sorting. For colonic whole mesenchyme sc-RNA-seq, cells were stained for EpCAM (eBioscience, 12-5791-83, 1:100) and CD45 (Invitrogen, 17045182, 1:100) by incubation for 25min at 4C; and cells were sorted for EpCAM- CD45-.  Single cell (sc)-RNAseq of PdgfraH2BeGFP cells proceeded on unstained cells sorted for GFP-level. 8,000 live (DAPI-negative) cells were loaded to a 10X Chromium controller proceeding with the Single Cell 3′ V3.1 assay (10X Genomics). Single Cell 3′ V3.1 assay (10X Genomics).