title: GSM4278223: SLX.14773.A011; Drosophila melanogaster; RNA-Seq
library_construction_protocol: 15 fly heads was homogenised by manual grinding in an eppendorf tube with 50μl Trizol (Cat No. 15596-018; Invitrogen). A further 50μl of Trizol were mixed with the homogenate, which was then microfuged at 16,160 x g for 10 minutes and the supernatant transferred to RNAse-free tubes before adding a further 50μl Trizol and incubated for 5 minutes. RNA was isolated by conventional chloroform:isoamylalcohol:isopropanol extraction followed by ethanol precipitation. Air-dried RNA pellets were treated with DNAse 1 (Cat No. EN0521; Thermo Scientific) at 37 ̊C for 60 minutes, with the reaction halted by the addition of EDTA and incubation at 65 ̊C for 10 minutes. RNA samples were then subjected to the Qiagen RNeasy RNA Cleanup protocol according to the manufacturers' instructions. The RNA samples were eventually eluted from RNeasy mini-spin columns, ethanol precipitated, re-suspended in RNAse-free water and stored at -80 ̊C. A library of mRNA was prepared from 200 ng of total RNA, extracted as described above and that had been originally pooled from 15 fly heads from each experimental condition, using the Illumina TruSeq Stranded mRNA Library Prep kit (Illumina, San Diego, CA, USA) according to the manufacturer's protocol.
identifiers.namespace: GEO
identifiers.id: GSM4278223