GSM8256113: Control, 2-week inducible, ScRNAseq; Mus musculus; RNA-Seq
Fresh back skin sample were clooected from control and Piezo1iKO C57BL/6 mice. The tissues were minced into small pieces and incubated in HBSS with 2.5 mg/ml collagenase I at 37 degress for 2 hours. Tissues and cells were collected by centrifuge and digested by 0.25% trypsin at 37 degree for 7 minutes. Single skin cells were washed by cold PBS. Library was performed according to the manufacter's instruction (Single cell 3' v3.1 protocol, 10x Genomics). Briefly, back skin cells were resuspended in the master mix and loaded together with gel beads and partitioning oil 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 retrotranscribed into cDNA. The pooled the cDNA was cleaned up with Silane DynaBeads, amplified by PCR and appropriate sized fragments were selected for subsequent library construction.