RNA-seq of Bombyx mori: 5th lavae fat body
A total RNA amount of 3 g per sample was used as input material for RNA sample preparations. First, ribosomal RNA was removed by Epicentre Ribo-zero rRNA Removal Kit (Epicentre, USA), and rRNA free residue was cleaned up by ethanol precipitation. Subsequently, sequencing libraries were generated using the rRNA-depleted RNA by NEBNext Ultra Directional RNA Library Prep Kit for Illumina (NEB, USA) following manufacturers recommendations. At last, products were purified, and library quality was assessed on the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumina) according to the manufacturers instructions. After cluster generation, the libraries were sequenced on an Illumina Hiseq 2500 platform and 125 bp paired-end reads were generated.