title: GSM4828849: F_Obp50b_4_63_S43; Drosophila melanogaster; RNA-Seq
library_construction_protocol: We collected replicates of 30 adult mated flies, sexes separately, of up to two lines per genotype between 8:00 am and 11:00 am and flash froze them on dry ice in 2 mL tough microtubes (Thermofisher, Waltham, MA). Total RNA was extracted using a modified version of the RNAeasy plus mini kit (Qiagen, Hilden, Germany) protocol. Four 2.4 mm metal beads (Thermofisher) were added to each sample tube and the flies were homogenized in a bead mill (Thermofisher) for 2 minutes at 5m/s, after which the RNA was eluted with a total of 30 μL H2O. We used the NuQuant +UDI, Drosophila AnyDeplete kit (Tecan, Männedorf, Switzerland) to deplete ribosomal RNA and prepare bar-coded cDNA libraries for sequencing after 17 cycles of amplification. We used the Qubit 1X HS dsDNA HS kit (Thermofisher) to quantify the libraries and high sensitivity D1000 screentape (Agilent, Santa Clara, CA) to check the quality of size selection. We then normalized the libraries to 5nM concentration and pooled them in order to get a final equimolar concentration of 3nM.