GSM5419507: DM T0 RNA 2; Drosophila melanogaster; RNA-Seq
150 larval brains per replica carrying GFP positive actin>F/O clones overexpressing UAS-HA-mγ, UAS-HA-dpn  (DM) were hand dissected, dissociated with collagenase and GFP positive cells were purified by FACS sorting and stored in Trizol. In parallel, larval brains of the same genotype were allografted into adult w1118 host flies and grown for 8-10 days. 25 T0 host flies were sacrificed per replica and the GFP tumour masses were collected in Trizol. Some tumour masses were used to generate more advanced allograft stages. In the forth allograft stage (T3), 25 host flies were again sacrificed per replica and GFP tumour masses were stored in Trizol.  RNA was extracted according to standard protocols. NGS libraries were generated using total RNA as input with polyA mRNA magnetic isolation kit (NEB) and the NEB Ultra II RNA library kit for Illumina kit according to manufactures protocol, using 13 cycles of amplification. Libraries were sequenced on Illumina Nextseq 500 on 1 x 75 High flowcell.