GSM6569817: WTFU; Mus musculus; RNA-Seq
After the sacrifice of wild-type and HrasG12V/+-mosaic models at 6 days post-tamoxifen injection (3 days post-wound induction), the uninjured and injured ears of each mouse were cut in small pieces with a punch biopsy of 8 mm in diameter (the wound was kept at the center the 8 mm biopsy to mostly isolate cells involved in injury-repair). The ear epidermis was dissociated from the dermis and incubated in 0.25 % Trypsin at 37°C for 30 minutes. The epidermal preparation was placed in a 70 μm cell strainer, smashed with a piston and rinsed three times with PBS + 0.04% BSA. The flow-through was subsequently filtered through a 40 μm cell strainer, spun-down and resuspended in 300-400 μl of PBS + 0.04% BSA. The viability of the cell suspension was determined using trypan blue. Cellular suspensions were counted and diluted to a final concentration of 1200 cells/μl in PBS/0.04% BSA and then loaded on a Chromium Controller to generate single-cell gel bead emulsions, targeting 3′. Libraries were sequenced on the Illumina Novaseq 6000 system in the following configuration: 28, 10, 10, 90 (read1, i7 index, i5 index, read2)