GSM9485403: Irf2KO; Mus musculus; RNA-Seq
Skin cells were isolated as described 46 with slight modifications. Mouse skin taken from the chest was digested with 1000 U/ml collagenase (Gibco, 17104019) and 20 U/ml DNase I (Sigma-Alrich, DN25) in cell dissociation buffer (ThermoFisher, 13150016) for 10 min at 37°C. Skin samples were digested two additional times, replenishing the digestion solution after spinning samples down. Tissue was mechanically disrupted by pipetting after each digestion. Single cell suspensions were filtered through 70 μm cell strainers and processed using the 10x Genomics Chromium Single Cell 3′ V3.1 Reagent Kit (PN-1000121). For each sample, we 39 targeted 10,000 viable cells. GEMs were generated, cells were lysed, and mRNAs were barcoded and underwent reverse transcription to produce full-length cDNAs. cDNAs were amplified with 12 PCR cycles. Gene expression libraries were sequenced at a depth of 20,000 reads/cell on an Illumina NovaSeq6000