GSM9699183: Aged POWV-infected brain (replicate 1); Mus musculus; RNA-Seq
Kit used: Miltenyi Brain Dissociation Kit Enzymatic treatment: Papain and DNase I Mechanical dissociation: gentleMACS Dissociator Incubation conditions: Time and temperature by default kit protocol Filtration: 70μm followed by 40μm cell strainers Libraries were prepared according to the manufacturer's instructions using the 10x Genomics Chromium Single Cell 3' v3.1 platform. Briefly, single-cell suspensions were loaded with barcoded gel beads into the Chromium Controller to generate gel bead-in-emulsion (GEM) partitions. Polyadenylated RNA from individual cells was reverse transcribed within GEMs to generate barcoded cDNA containing unique molecular identifiers (UMIs) and cell barcodes. Following reverse transcription, GEMs were broken, and cDNA was purified, amplified by PCR, and subjected to fragmentation and size selection. Sequencing adapters and sample indices were added during library construction to generate paired-end sequencing libraries compatible with Illumina platforms.