GSM8897925: Middle phalanx (P2) cells, BSA treated; Mus musculus; RNA-Seq
Cell collection was carried out by removing the surrounding epithelium from the amputated digits and dissecting the tissue distal to the P2 stump, followed by placing the tissue into ice-cold PBS. Enzymatic dissociation was perform using 2U/ml Liberase Blendzyme (37°C, 4h; Sigma-Aldrich Co.), with manual pipet trituration every 2 hours. Cells were filtered to remove aggregates, washed and resuspended in 0.8% BSA in PBS. Single cells were captured in Gel Beads-in-Emulsion (GEMs) using the Chromium Controller with Chromium Next GEM Single Cell 3' Reagent Kits v3.1 (10X Genomics, Pleasanton, CA). Barcoded cDNA was generated through reverse transcription, followed by amplification and fragmentation. Dual-indexed sequencing libraries were constructed according to the 10X Genomics Single Cell 3' v3.1 protocol.