title: GSM5957302: Alk-YS, 2; Drosophila melanogaster; RNA-Seq
library_construction_protocol: 3rd instar larvae (40 - 45) were collected and quickly washed in water to remove food leftovers and yeast. Larvae were placed in ice-cold PBS of a depression well. Fine forceps were used to dissect the larval CNS. Once the larval CNS was exposed, it was dissected out using forceps, and the intact CNS were collected in an Eppendorf tube containing 250 μl of ice-cold solution (equal volume of both RNA free PBS and RNALater solution from Invitrogen) for a maximum of one hour. Afterwards, the tubes containing the CNS were centrifuged at 2000 rpm for 5 minutes at 4 °C and removed the supernatant. 500 μl fresh RNALater solution was added to the CNS and were stored at -80°C. Once the CNS collection for 3 biological replicates per genotype was obtained, RNA-extraction was carried out according to the manufacturer's protocol (Promega ReliaPrepTM RNA Tissue Miniprep System, REF-Z6111). Total RNA was measured using NanoDrop OneC (Thermo Scientific) for its concentration and RNA-integrity was checked on the gel electrophoresis. 4-6 µg of total RNA/biological replicate was shipped to Novogene Co. Ltd (UK) for sequencing. cDNA