title: GSM6735727: Control_rep1; Mus musculus; RNA-Seq
library_construction_protocol: The V-SVZ of the lateral walls of the lateral ventricles from one-month old (P31-33) mice were dissected according to published protocols (Mirzadeh et al., 2010). Single cell suspensions were prepared as described in (Brewer and Torricelli, 2007). Briefly, a 2 mg/ml papain solution was pre-activated at 37℃ for 20 min. The V-SVZ samples were then digested in the papain solution at 30℃ for 30 min with a shaking speed of 60 rpm in a shaker. The tissue sample is then mechanically dissociated by trituration with a Pasteur pipette 10~15 times per round for three rounds, washed, and centrifuged with an OptiPrep density gradient to remove myelin and cellular debris. After centrifugation, the dense white layer and all layers above were carefully aspirated. The lower layers were pelleted at 200g for 3min, resuspended in 1 ml PBS with 0.04% bovine serum albumin (0.04% BSA-PBS), filtered through 40 μm strainers, and resuspended in 0.04% BSA-PBS at 700-1,200 cells/µl for single cell capture. Single cell gene expression libraries were created using Chromium Next GEM Single Cell 3' (v3.1 Chemistry) (10X Genomics), Chromium Next GEM Chip G Single Cell Kit (10X Genomics), and Single Index Kit T Set A (10X Genomics) according to the manufacturer's instructions. Cells were loaded to target 10K cells to form GEMs and barcode individual cells. GEMs were then cleaned, and cDNA libraries were created according to manufacturer's instructions. Library quality was assessed using 4200 TapeStation System and D1000 ScreenTape (both from Agilent) and Qubit 2.0 (Invitrogen) for concentration and size distribution. 