GSM2491833: Dmel_72h_repC; Drosophila melanogaster; RNA-Seq
Larvae were dissected and eye-antennal imaginal discs were stored in RNALater at 72/96/120h after egg laying. 40-50 discs were dissected for the 120h samples, 80-90 discs for the 96h samples and 120-130 discs for the 72h samples. Total RNA was isolated using Trizol  and the samples were DNase I treated in order to remove DNA contamination. Library preparation for RNA-Seq was performed using the TruSeq RNA Sample Preparation Kit starting from 500 ng of total RNA. Accurate quantitation of cDNA libraries was performed by using the QuantiFluor™dsDNA System . The size range of final cDNA libraries was determined applying the DNA 1000 chip on the Bioanalyzer 2100 from Agilent (280 bp). cDNA libraries were amplified and sequenced (50 bp single-end reads) using cBot and HiSeq 2000 (Illumina). Sequence images were transformed to bcl files using the software BaseCaller (Illumina). The bcl files were demultiplexed to fastq files with CASAVA (version 1.8.2).