title: GSM4875738: Control rep 1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: 40 brains per sample at the mid-3rd instar stage were dissected in cold AHL (108 mM NaCl, 5 mM KCl, 2 mM CaCl2, 8.2 mM MgCl2, 4mM NaHCO3, 1 mM NaH2PO4-H2O, 5 mM trehalose, 10 mM sucrose, 5 mM HEPES). Three biological replicates for the control group and four for the experimental, each with 40 brains, were dissected. Brains were transferred to 1 mL of Trizol and incubated for 5 minutes at room temperature (RT). 0.2 mL of chloroform was added and samples were inverted. Samples were incubated 2-3 minutes at RT, then centrifuged at 12,000g for 15 minutes at 4oC. Genomic DNA was removed using a gDNA eliminator column (RNeasy Plus Micro Kit, Qiagen). RNA was then extracted using the RNeasy MinElute Cleanup Kit (Qiagen). Sequence libraries for each sample were synthesized using the NEBNext Ultra II Directional RNA kit following supplier recommendations and were sequenced on Illumina HiSeq-4000 sequencer as single reads of 100 base reads following Illumina's instructions.