title: RNA-seq of Daphnia magna animals at age of 10 days exposed to Zinc sub-lethal concentrations
design: RNA was isolated using the Promega SV Total RNA Isolation System according to the protocol supplied by the manufacturer. The quantity of the isolated RNA was determined using a Thermo Scientific NanoDrop 2000 spectrophotometer and a Invitrogen Qubit 2.0 Fluorometer. Integrity of the isolated RNA was evaluated by gel-electrophoresis and by using a Aligent 2100 Bioanzalyer using an Eukaryote Total RNA Nano chip. Per treatment 4 samples  were selected and prepared for sequencing using the Illumina TruSeq RNA Library Preparation Kit v2 according to the protocol supplied by the manufacturer. Briefly in this procedure, total RNA is purified for mRNA using oligo-dT attached magnetic beads and, subsequently, mRNA is fragmented. Hereafter, mRNA is transformed into cDNA via reverse transcriptase and sample unique adapters are ligated to the cDNA fragments. Quality and quantity was assessed using an Aligent 2100 Bioanalyzer using a DNA 7500 chip. After additional clean-up procedure, samples were diluted to 10 mol/l cDNA and subsequently pooled. Samples were sequenced at the VUmc Amsterdam using Illumina HiSeq 2500.