title: Illumina HiSeq 4000 paired end sequencing; RNA-seq on Drosophila wing imaginal discs after Myc RNAi or overexpression
design: RNA-seq on Drosophila wing imaginal discs after Myc RNAi or overexpression
library_construction_protocol: For each replicate about 60 wing discs were dissected from wandering larvae and immediately transferred to the lysis buffer (Buffer RLT, Qiagen) supplemented with β-mercaptoethanol (1:100) and stored at -80°C until further processing.   Eggs were collected in bottles for 10h at 18°C, and then incubated for 4 days at 18°C (permissive temperature for Gal80TS), and finally transferred for 2 days to 29°C (restrictive temperature for Gal80TS) to induce expression of the UAS transgenes.  Conditional expression of UAS constructs in the wing pouch was achieved by using the nub-Gal4 and the TARGET system. Total RNA from each sample was isolated using the RNasey mini kit (Qiagen) according to manufacturer's instruction and subsequently quantified with the Qubit 3.0 Fluorometer and the Qubit RNA BR Assay Kit (Thermo Fisher).  RNA-seq libraries were generated with the Ovation Drosophila RNA-Seq System (NuGEN) which requires 10 and 100 ng of input RNA.