GSM5236530: mouse_E85_spinalcord_rep1; Mus musculus; RNA-Seq
Dissected samples were snipped into smaller pieces and incubated on a FACSmax (Amsbio, T200100) cell dissociation solution containing 10X Papain (30 U/mg, Sigma-Aldrich, 10108014001) for 11 min at 37°C to dissociate the cells and transferred to a HBSS solution with 5% FBS, Rock inhibitor (10 μM, Stemcell Technologies, Y-27632) and 1X of non-essential aminoacids (Thermo Fisher Scientific, 11140035). Single cells were disaggregated through pipetting and filtered, and quality control was assayed by measuring live cells versus cell death, cell size and number of clumps on an EVE cell counter. Each sample was prepared at a concentration of 600-1700 cells/µl and viability ranged between 70-96%. Samples with a viability above 70% were used for sequencing. RNA libraries were prepared for sequencing using Chromium Single Cell 3' v2; cDNA amplification involved 12 PCR cycles