GSM8462151: Old_WT_02; Mus musculus; RNA-Seq
RNA was isolated using the Invitrogen purelink RNA micro kit (cat#12183-016) and processed as per manufacturer's instructions. Single-cell libraries were prepared in triplicate with >85% cell viability confirmed by trypan blue staining. Cells were resuspended in MEM with 10% FBS at 1200 cells/uL. Library construction used Chromium Next GEM Single Cell 3ʹ dual index Kits v3.1 with Chip G (10x Genomics). About 10,000 single cells were loaded to generate gel bead-in-emulsions (GEMs), lysed, and RNA was barcoded and reverse-transcribed. GEMs were broken, and cDNA was prepared following the manufacturer's protocol. cDNA quality was assessed with the Agilent Bioanalyzer and Pico-green. Libraries underwent QC by shallow-sequencing on an Illumina NextSeq and final sequencing on an Illumina Nova Seq S4 capturing ~20,000 paired-end reads per cell. Sequencing libraries were converted into feature-barcode matrices using Cell Ranger (10x Genomics, version 5.0.0) and mapped to GRCh38.