GSM1666107: L2 rep1; Drosophila melanogaster; RNA-Seq
For tissue dissociation, pupal brain tissue dissected at 40hrs after pupal formation was incubated with a Papain (Worthington) and Liberase TM protease cocktail (Roche) at 250C for 15 min in a microfuge shaker at 1000rpm. At 5 and 10 min into this incubation, the tissue was pipetted up and down with a P200. At 15 min the sample was passed through a 25G 5/8-gauge needle until the tissue was completely dissociated. Digestion was inactivated by the addition of rich media with serum, and the cell suspension was passed through a 70μm filter. To concentrate the cells, the suspension was spun down at 1600rpm for 8 min at 40C. After decanting the supernatant, cells were re-suspended in approximately 500ul of rich media and sorted in a BD FACSAria II. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM1666107