Truseq pe cluster kits v3 cbot hs

Total RNA from the silkworm's fat body tissue at 2 and 6 h.p.i.20E was extracted using the Trizol reagent (Invitrogen) and further purified with RNeasy kits (Qiagen).
RNA purity was checked using a NanoPhotometer spectrophotometer (IMPLEN, CA, USA). RNA concentration was measured using Qubit RNA Assay Kits in a Qubit 2.0
Flurometer (Life Technologies, CA, USA). RNA integrity was assessed using RNA Nano6000 Assay Kits from the Bioanalyzer 2100 system (Agilent Technologies, CA, USA).
A total of 3 μg RNA per sample was used as input material for RNA sample preparation. Ribosomal RNA was removed using Epicentre Ribo-zero™ rRNA Removal Kits (Epicentre, USA), and rRNA-free residue was cleaned up by ethanol precipitation. Subsequently, sequencing libraries were generated using the rRNA-depleted RNA with NEBNext Ultra™ Directional RNA Library Prep Kits for Illumina (NEB, USA), following manufacturer's recommendations. Finally, the products were purified, and the library quality assessed using the Agilent Bioanalyzer 2100 system. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kits v3-cBot-HS (Illumina), according to the manufacturer's instructions. After cluster generation, the libraries were sequenced on an Illumina HiSeq 2500 platform and 125 bp paired-end reads were generated.