title: GSM9409765: C2 rep1; Bos taurus; RNA-Seq
library_construction_protocol: nuclei were extracted from snap frozen cells using Miltenyi tissue dissociator Isolated nuclei (0.7–1.2 million nuclei/mL, 200 µL) were submitted to the University of Missouri Genomics Technology Core. Nuclei concentration and viability were assessed using acridine orange/propidium iodide staining and a Cellometer K2 (Nexcelom). Approximately 10,000 nuclei per sample were processed following the 10x Genomics Chromium Next GEM Single Cell Gene Expression protocol. Libraries were generated by cDNA synthesis, fragmentation, end repair, A-tailing, adapter ligation, barcoding, and clean-up as recommended by the manufacturer. The resulting libraries were quantified, size-assessed, pooled, and sequenced on an Illumina NovaSeq X+ instrument, targeting 50,000 reads per nucleus with a sequencing configuration of 28 bp on Read 1 and 100 bp on Read 2. Raw sequencing data were then delivered for downstream analysis.Single-nuclei RNA-seq analysis.