GSM8866299: JiaWT, scRNAseq; Mus musculus; RNA-Seq
Neocortices were incubated with a prewarmed solution of Papain (Worthington; prepared according to manufacturer's protocol) for 15 minutes at 37°C. After Papain incubation, tissues were washed multiple times with DMEM/10% FBS (Thermo Fisher) and were then gentle triturated. The tissue was allowed to settle for about 1 minute. The cell-containing supernatant was filtered (30 μm MACS SmartStrainer) into new conical tubes. Conical tubes containing cells were centrifuged at 200 rcf for 2–3 minutes. The supernatant was removed without disturbing the cell pellets. The pellets were resuspended in cold Hibernate buffer (BrainBits) to achieve a concentration of 2,000 cells per microliter. Library preparation was performed by the UCSF Institute of Human Genetics Core (IHG), using 10X Chromium Single 3' Reagent Kits Version 2 (10X Genomics). The library length and concentration were quantified using a Bioanalyzer (Agilent) according to the manufacturer's protocol. Paired-end sequencing was performed on the Illumina NovaSeq at Center for Advanced Technology (UCSF).