title: GSM9035691: Sample11_scRNAseq_White_leghorn_contour_repeat; Gallus gallus; RNA-Seq
library_construction_protocol: Regenerating feather follicles were surgically dissected and only 0.5 cm feather follicles with DP and DS were used for scRNA-seq (Fig. 4A). Feather follicles were cut to small pieces and incubated in 5 ml RPMI solution containing 232U DNAse I (Sigma-Aldrich), 1.25mg Liberase (Sigma-Aldrich), 5mg dispase:collagenase (Sigma-Aldrich) for 2 hours at 37°C and quenched with chelated FBS. Cells were passed through a 70-µm filter and centrifuged at 500g for 5 min, and the pellet was resuspended in PBS free of Ca2+ and Mg2+and 1% BSA. Dead cells were removed by using Dead Cell Removal Kit (Miltenyi Biotec). Live cells were resuspended in 0.04% UltraPure BSA (Sigma-Aldrich) and counted using the automated cell counter Countess (Thermo Fisher Scientific). Cells were captured using Chromium (10X Genomics). GEM (Gel Bead-In EMulsions) generation, barcoding, post–GEM-RT (reverse transcription) cleanup, complementary DNA (cDNA) amplification, and cDNA library construction were performed using Single-Cell 3′ v2 chemistry (10X Genomics).