GSM9065010: WT; Drosophila melanogaster; RNA-Seq
Single-nucleus suspension was conducted as described by Li et 2022. Briefly 70 guts per sample were dissected in cold Schneider's medium, flash-frozen and stored at -80°C. Prior to FACs sorting, samples were spined down and Schneider's medium was exchanged with homogenization buffer (Li et 2022). Using 1ml dounce , nuclei were released by 20 loose pestle strokes and 40 tight pestle strokes while keeping samples on ice and avoiding foam. Next, nuclei were filtered through 5 ml cell strainer (40 μm), and using 40 μm Flowmi. Nuclei were centrifuged,  resuspended in PBS/BSA with 0.5% RNase inhibitor and filtered again with 40 μm Flowmi. Single nuclei were sorted using Sony SH800Z Cell Sorter and 100k nuclei per sample were collected in PBS/BSA buffer. Libraries were generated according to 10X genomics Next GEM Single Cell 3' Library and Gel Bead Kit v3.1