GSM7755603: ALG, TDP43 overexpression, replicate 2; Drosophila melanogaster; RNA-Seq
Two day old female flies were anesthetized with CO2 and flash frozen in liquid nitrogen before being stored at -80°C until an appropriate number of flies were collected for each group (>1,000,000 cells for all groups except SPG, for which cell number was >300,000). Fly heads were separated, homogenized, and incubated with an antibody against GFP before being bound to protein A beads, cleaved, and rebound to protein G beads. Twice purified TAPIN-tagged nuclei were then burst and nuclear RNA was collected and stored at -80°C TAPIN-generated nuclear RNA was treated with DNase and purified by the Arcturus PicoPure system. Purified RNA was amplified to cDNA via Nugen Ovation v2 system and was then fragmented with a Covaris s-series to 200bp. Fragmented cDNA was repaired and underwent second strand synthesis before being ligated to barcoded linkers.