GSM4676969: P30 Braf WT rep 2; Mus musculus; RNA-Seq
Procedure was conducted at either post natal day 30 (P30) or post natal day 50 (P50). Mice were euthanized at the appropriate age and the hair on the dorasal skin was removed by shaving and depilation. A 2x3 cm section of the dorsal skin was removed, and the fat scraped off from the underside. The piece was then diced into smaller pieces and suspended in dissociation buffer (RPMI, liberase, Hepes, Sodium Pyruvate, Collagenase:Dispase) for 50 minutes at 37°C with gentle agitation. After incubation, DNaseI was added for 10 minutes and then inactivated with fetal bovine serum and EDTA.  The tissue suspension was further dissociated mechanically with the GentleMACS.  Single cell suspensions were filtered twice through a 70µm strainer and dead cells removed by centrifugation.  The live cells were washed with 0.04% UltraPure BSA:PBS buffer, gently re-suspended in the same buffer. Samples were loaded onto the 10X Chromium platform. Libraries were prepared using the Chromium Single Cell 3' v2 protocol (10X Genomics).  Briefly, individual cells and gel beads were encapsulated in oil droplets where cells were lysed and mRNA was reverse transcribed to 10X barcoded cDNA.  Adapters were ligated to the cDNA followed by the addition of the sample index.  Prepared libraries were sequenced using paired end 100 cycles chemistry for the Illumina HiSeq 4000.
GEO Accession
GSM4676969