GSM8807704: PKI_2; Mus musculus; RNA-Seq
Keratinocytes were isolated from BCC mice and controls 28 days post-tumor induction. Tail skin was removed from mice, cleaned using 10% iodine in PBS and washed with sterile PBS several times. Each tail was cut into pieces and then incubated overnight at 4°C in 2.5 parts of 0.25% trypsin (Gibco 15050-057) and 1 part of 5U/mL dispase (Stem Cell Technologies 07913). The next day, the epidermis was scraped from the dermis and minced in 0.25% trypsin with EDTA (Gibco 25200056). Trypsin was neutralized in DMEM with 10% FBS and cells were strained in a 100μm cell strainer followed by a 40μm cell strainer. After centrifugation at 500G for 8 minutes, cells were collected in EpiLife media and processed for single-cell RNAseq. The cellular samples were loaded in the lanes according to the 10X Genomics Chromium Next GEM Single Cell 3'v3.1 (Dual Index) User Guide with a single capture lane per sample. All subsequent steps of library preparation and quality control were performed as described in the user guide on the Chromium Connect. Two NovaSeq runs for GEX were performed (indicated as Seq1 and Seq2).