GSM3352706: E13.1H_SCHT6_5_14; Mus musculus; RNA-Seq
The putative primary somatosensory cortex S1 was microdissected and incubated in 0.05% trypsin at 37°C for 5 minutes. Following tissue digestion, fetal bovine serum was added to the mix and cells were manually dissociated via up-and-down pipetting. Cells were centrifuged 5min at 300g and the pellet was suspended in 1ml of HBSS then passed on a 70µm cell stainer. FT+ cells, gated on the top 5% brightest cells, were finally FAC-sorted on a MoFloAstrios (Beckman). FAC-sorted FT+ cells (18µl) were mixed with the C1 Suspension Reagent (2µl; Fluidigm) yielding a total of 20µl of cell suspension mix with ~500 cells/μl. The cell suspension mix was loaded on a C1 Single-Cell AutoPrep integrated fluidic circuit (IFC) designed for 10- to 17-µm cells (HT-800, Fluidigm #100-57-80). cDNA synthesis and preamplification was processed following the manufacturer's instructions (C1 system, Fluidigm). Single cell RNA-sequencing libraries of the cDNA were prepared using Nextera XT DNA library prep kit (Illumina). Libraries were multi-plexed and sequenced according to the manufacturer's recommendations with paired-end reads using HiSeq2500 platform (Illumina) with an expected depth of 1M reads per single cell, and a final mapping read length of 70bp. SMARTseq v4 kit (Clontech, # 634888)
GEO Accession
GSM3352706