title: GSM9240887: LVFW_WT_13mo; Sus scrofa; RNA-Seq
library_construction_protocol: Porcine cardiac tissue samples were processed by an experimenter blinded to genotype. Nuclei were isolated from unfixed frozen tissue using detergent-based lysis, purified by fluorescence-activated cell sorting, and microscopically inspected to confirm integrity and purity. Nuclei were quantified with a Countess II system (Life Technologies), kept on ice, and immediately used for single-nucleus RNA-sequencing Single-nucleus suspensions were loaded onto a Chromium Controller (10x Genomics) targeting a recovery of 5,000–10,000 nuclei per reaction. 3′ gene expression libraries were prepared using the Chromium Single Cell 3′ Reagent Kits v3 (10x Genomics) following the manufacturer's instructions. cDNA quality was assessed with the Agilent Bioanalyzer High Sensitivity DNA Analysis Kit, and libraries were quantified using the KAPA Library Quantification Kit. Sequencing was performed on an Illumina NovaSeq 6000 platform at a target depth of 30,000–50,000 reads per nucleus.