GSM8972283: male, central brain dissection, 5-day-old, replicate 2; Drosophila melanogaster; RNA-Seq
2 replicates each of 20 male and 20 female central brains were individually dissected, removing optic lobes, in toxin-supplemented ice-cold calcium- and magnesium-free DPBS (GibcoTM, + 50 mM D(−)-2-Amino-5-phosphonovaleric acid, 20 mM 6,7-dinitroquinoxaline-2,3-dione and 0.1 mM tetrodotoxin). Each replicate was then washed in 1 mL ice-cold toxin-supplemented Schneider's medium (tSM: GibcoTM + toxins, as above). Brains were then incubated for 30 minutes in 0.5 mL of tSM containing 1 mg/mL papain (Sigma-Aldrich) and 1 mg/mL collagenase I (Sigma-Aldrich). Brains were washed once more with tSM and subsequently triturated with flame-rounded 200 mL pipette tips. Dissociated brains were resuspended into 1 mL PBS + 0.01% BSA and filtered through a 10 µm CellTrics strainer (Sysmex, 04-0042-2314). Libraries were made using the Chromium Single Cell 3' v2 kit from 10x Genomics. Cells were loaded in accordance with 10x Genomics documentation. The samples were sequenced with 8 lanes of Illumina HiSeq4000 by the Oxford Genomics Centre.
GEO
GSM8972283_r1