GSM7436622: P21-28_ctrl_Sst68; Mus musculus; RNA-Seq
At postnatal day 21-28 (P21-28) the animals were anesthetized with ketamine/xylazine (4:1), then transcardially perfused with cutting solution of the following composition (in mM): 87 NaCl, 75 sucrose, 2.5 KCl, 25 NaHCO3, 1.25 NaH2PO4, 7 MgCl2, 1 CaCl2, and 10 glucose. The brains were quickly removed and transferred to ice-cold cutting-solution, subsequently fixed to a stage and 300 μm brain slices were cut on a vibratome (VT1200 S, Leica). Tissue corresponding to primary somatosensory cortex (S1) was dissected and dissociated into a single cell suspension, as previously described (Muñoz-Manchado et al. 2018; DOI: 10.1016/j.celrep.2018.07.053). Briefly, the tissue was dissociated using the Papain dissociation system (Worthington) following the manufacturer's instructions. Cell suspension obtained was filtered with 20 μm filter (Partec) and kept in cold oxygenated Hank's balanced buffer solution (HBSS; Sigma) with 0.2% bovine serum albumin (BSA) and 0.3% glucose. Cells were sorted based on fluorescence (eGFP or tdTomato), and then loaded on the Fluidigm C1 platform, where they were processed according to the C1-STRT protocol (Zeisel et al. 2015;  DOI: 10.1126/science.aaa1934); including imaging, high-throughput sequencing (Illumina) and molecule counting (UMI). Visual doublets were excluded, and only fluorescent single cells were processed for RNA sequencing. The libraries were generated according to the C1-STRT protocol (Islam et al., 2014; DOI: 10.1038/nmeth.2772).