midgut transcriptome of Plateumaris braccata
midguts were extracted, snap-frozen in liquid nitrogen and stored at -80 C. Total RNA was extracted using the innuPrep DNA/RNA Mini kit (Analytik Jena, Jena, Germany) following the manufacturer's instructions. Genomic DNA contamination was removed by DNAse treatment (TURBO DNase, Invitrogen, Carlsbad, CAL, USA) for 30 min at 37 C. Total RNA samples were further purified by using the RNeasy MinElute Clean up Kit (Qiagen, Hilden, Germany) following the manufacturers protocol, except for elution to 20 l volume. The integrity and quality of the RNA samples were determined using the RNA 6000 Nano LabChip kit on an Agilent 2100 Bioanalyzer (both Agilent Technologies, Santa Clara, CAL, USA) according to the manufacturers instructions. RNA-Seq was outsourced to the Max Planck Genome center (Cologne, Germany). There, poly(A)+ enriched RNA was fragmented to an average of 300-350 nucleotides and a TruSeq compatible, directional library was prepared for each sample. Libraries were tagged using dual indexing and were multiplexed on the same sequencing lane. Sequencing was carried out on a HiSeq3000 sequencing platform (Illumina, CA, USA) using paired-end (2 x 150 bp) reads.