title: GSM3874364: S1_FJ_Wild type; Mus musculus; RNA-Seq
library_construction_protocol: Tissue harvesting: Embryos were harvested at E15.5 then washed in ice cold HBSS. Trunk skin was then removed, rinsed in HBSS and digested in 300ul of 0.2 U/ml dispase and 0.5mg/ml DnaseI in HBSS for 45 minutes at 37 degrees celcius. Digestion was halted by the addition of 5mM EDTA/PBS. dissociation protocol: Skin suspensions were dissociated by passing through 18G then 21G needles, then filtered through 50 um filter membranes. Cell suspensions were centrifuged at 300g for 5 minutes at 4 degrees celcius, before being resuspended in sorting buffer (1% FBS, 5mM EDTA and 25mM HEPES in PBS). cell sorting protocol: tdTomato negative skins were used as a negative control for gating and 1 ug/ml DAPI was used as a dead cell marker prior to sorting. TdTomato positive live cells were sorted using a BD FACSAria cell sorter. Approximately 7000-10000 cells per sample were used for single cell RNA isolation, cDNA synthesis and library preparation using a 10X Genomics Chromium Single Cell 3' Reagent kit as per the manufacturer's protocol. Libraries were sequenced using an Illumina HiSeq4000 sequencer.
attributes.tag: GEO Accession
attributes.value: GSM3874364