RNA-seq of Drosophila melanogaster: third instar larval brain
The RNA-Seq of each individual sample was carried out from IGA Technology Services (Udine, Italy). cDNA libraries were constructed with 1 g of total RNA by using the "Ovation Drosophila RNA-Seq System'' (TECAN-NuGEN, Redwood City, CA, USA) following the manufacturer's instructions. The total RNA was fragmented 3 min at 94 C. 1X Agencourt AMPure XP beads (Agencourt Bioscience Cooperation, Beckman Coulter, Beverly, MA, USA). Total RNA samples and final cDNA libraries were quantified with the Qubit 2.0 Fluorometer (Invitrogen, Carlsbad, CA, USA) and quality tested by Agilent 2100 Bioanalyzer Nano assay. For cluster generation on the flow cell, libraries were processed with cBot (Illumina, San Diego, CA, USA) following the manufacturer's instructions. Sequencing was carried out in single-end mode (175 bp) by using NextSeq500 (Illumina) with a targeted sequencing depth of about 40 million reads per sample. Raw data were processed with the software CASAVA v1.8.2 (Illumina) for both format conversion and de-multiplexing.
SUB10002092
6B-B