title: GSM9170448: Normal_rep 3; Mus musculus; RNA-Seq
library_construction_protocol: Adult male FVB mice were anesthetized with 1.25 % tribromoethanol i.p. and transcardially perfused with ice-cold artificial cerebrospinal fluid (ACSF). Cortices were micro-dissected in cold ACSF, stripped of meninges and white matter, then dissociated into single-cell suspensions using the Papain Dissociation System (Worthington Biochemical, #LK003176) following the manufacturer's protocol. Viable cells (>85 %) were counted by trypan blue exclusion and kept on ice for downstream processing. Single-cell suspensions (2-2.5 × 10^5 cells ml⁻¹ in ice-cold 0.04 % BSA/PBS) were loaded onto a 150 000-well SCOPE-chip™ microfluidic device (Scope Single-Cell Technologies). Gravity-driven Poisson loading yielded ≈7 000 single cells. Barcoded magnetic beads carrying cell-specific oligos and UMIs were introduced (one bead per well). After on-chip lysis, poly-adenylated mRNA was captured by oligo(dT) beads. Beads were pooled and reverse-transcribed using SuperScript IV (Thermo Fisher) followed by 12–14 cycles of PCR amplification (KAPA HiFi HotStart). Amplified cDNA (average 350 bp after tagmentation) was processed for tagmentation, end-repair, A-tailing and Illumina adapter ligation using the Nextera XT kit (Illumina). Libraries were quantified by Bioanalyzer (Agilent) and qPCR (KAPA Library Quantification kit) before sequencing.
identifiers.namespace: GEO
identifiers.id: GSM9170448_r1