GSM9059726: ipsilateral dorsal striatum, M3367 & F3325, Late learning; Mus musculus; RNA-Seq
Mice were anesthetized and perfused with cold, oxygenated ACSF. Coronal brain slices (300 µm) were prepared using a vibratome and incubated in ACSF at 3 °C for 30 min. Motor cortex (Fos-TRAP) and dorsal striatum (Arc-TRAP) were dissected, transferred to oxygenated HBSS/s solution, and pooled by hemisphere from two mice per condition. Tissue was digested with papain (20 U/ml, 37 °C, 1 h), triturated, and filtered through 40 µm strainers. Dead cells were removed using a Dead Cell Removal kit, and single cells were resuspended in cold PBS at 700–1200 cells/ml for downstream processing. cDNA libraries were prepared using Chromium Next GEM Single Cell 3' Library Construction Kit v2 or v3 (10x Genomics). Single-cell suspensions were loaded into the Chromium Chip for target recovery of 10,000 cells. cells were barcoded and lysed in droplets. Single-stranded cDNAs were then released from droplets, PCR amplified (11 cycles), fragmented, size-selected, ligated with adapters, and amplified by index PCR (11~12 cycles) to construct the final cDNA libraries.