GSM9108712: wild type sample3; Drosophila melanogaster; RNA-Seq
For each sample, 16 wandering third instar larvae (8 male, 8 female) that had not everted their spiracles were collected, washed with PBS, snap-frozen in liquid nitrogen, and stored at −80°C. Frozen larvae were homogenized in a glass dounce. Nuclei were released and lysate containing nuclei was filtered sequentially through cell strainers. The nuclei sample was filtered and washed, labeled with 1µM DRAQ7 Dye, and 500,000 healthy nuclei were isolated by a Sony SH800Z Cell Sorter. FACS sorted nuclei were adjusted to a final concentration of 1,000 nuclei/µL in 1xPBS/1%BSA. Libraries were generated according to 10X genomics Next GEM Single Cell 3' Library and Gel Bead Kit v3.1