GSM8606757: Back_Pdgfra+Krt5lineage-; Mus musculus; RNA-Seq
Freshly collected skins were minced using scissors, and suspended  in digestion buffer (RPMI with 5%v/v  fetal bovine serum, 0.3mg/mL Liberase TM (Merck) and 5U/mL DNase1) and incubated 37°C for 30minutes twice. The samples were resuspended in another digestion buffer (TrypLE Select Enzyme(Thermo Fisher Scientific) with 5U/mL DNase1) and incubated 37°C for 15minutes. The obtained cell suspensions were filtered through a 70µm cell strainer, washed in Minimum Essential Medium alpha (Thermo Fisher Scientific) with 2%v/v FBS. Isolated skin cells were individually labeled using the 3' CellPlex Kit Set A (10x Genomics). SYTOX negative target cells from each individual were sorted using the BD FACS Aria III system (BD Biosciences).Library preparation was performed according to the 10x Genomics protocol "Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (Dual Index) with Feature Barcode technology for Cell Multiplexing".