title: GSM8853763: WT, skin, scRNAseq; Mus musculus; RNA-Seq
library_construction_protocol: Skin collected from the flanks fof WT and TSK mice were pooled and minced into 1mm pieces. Tissue was further dissociated by enzymatic digestion by rotating at 37°C for 60 minutes with 10 mg/ml Collagenase I, 5 mg/ml collaganase IV, 10 mg/ml Collagenase D,and 5mg/ml DNAse I in DPBS. After dissociation, the suspension was strained (70 micron strainer) followed by centrifugation (8 minutes at 300g). Red blood lysis buffer (Thermo Fisher) was used followed by dead cell removal (STEMCELL) to deplete the samples of erythrocytes and dead cells, respectively according to the manufacturer's instructions. Finally, cells were resuspended in 0.04% BSA-PBS solution at a concentration of 700-1000 cells/microliter without selection for specific cell types. Library construction was performed according to the manufacturer's instructions (Next GEM 3' Gene Expression v3, 10x Genomics). Briefly, GCs were resuspended in the master mix and loaded together with partitioning oil and gel beads into the chip to generate the gel bead-in-emulsion (GEM). The poly-A RNA from the cell lysate contained in every single GEM was retrotranscripted to cDNA, which contains an Ilumina R1 primer sequence, Unique Molecular Identifier (UMI), and the 10x Barcode. The pooled barcoded cDNA was then cleaned up with Silane DynaBeads, amplified by PCR, and the appropriate-sized fragments were selected with SPRIselect reagent for subsequent library construction. During the library construction Ilumina R2 primer sequence, single-end constructs were added