GSM9555083: Control_30_female_body; Drosophila melanogaster; RNA-Seq
Nuclei were isolated following a previously published protocol (McLaughlin et al., 2022). Hoechst-stained nuclei—including polyploid populations—were purified using a BD FACSAria III cell sorter. For each 10x Genomics run, approximately 150,000–200,000 nuclei were sorted and pelleted by centrifugation at 1,000 × g for 10 min at 4 °C. The pellet was resuspended in 80 µL of 1× PBS supplemented with 0.5% BSA and RNase inhibitors. Nuclei were quantified using a hemocytometer, and 45,000–55,000 nuclei were loaded to recover ~20,000 nuclei per sample. Single-nucleus 3′ gene expression libraries were prepared with the Chromium Next GEM Single Cell 3′ HT Reagent Kits v3.1 (Dual Index) according to the manufacturer's protocol (10x Genomics). Libraries were sequenced on an Illumina NovaSeq X Plus platform (paired-end 150 bp; Novogene Co., Ltd.) to a depth of ~30,000–50,000 reads per nucleus. snRNA-seq