title: Boundary caps: a major source of skin mural cells
design: Boundary caps: a major source of skin mural cells
library_construction_protocol: Embryos at E12.5 were selected on the basis of Tomato expression under fluorescent stereomicroscope and head and viscera were removed.  The back skin was dissected from Krox20Cre/+,R26RTom/+ embryos, and then digested with collagenase/dispase type I (Merck/Roche) for 15 min at 37°C. Skin samples were then mechanically dissociated and the cell suspension was filtered. Dissociated cells were then resuspended in PBS, 1% BSA. Tomato-positive cells were isolated from embryonic skin cell preparations by FACS. Dead cells and doublets were excluded by gating on a forward-scatter and side-scatter area versus width. Tomato-positive cells were isolated by FACS from embryonic skin at E12.5. Around 10,000 cells were loaded into one channel of the Chromium system using the V3 single cell reagent kit (10X Genomics)  Following capture and lysis, cDNA was synthesized, then amplified by PCR for 12 cycles as per the manufacturer's protocol (10X Genomics).