RNA-Seq of Pagrus auratus gill tissue
Frozen tissue was partially thawed and submerged in lysis buffer containing 1% -mercaptoethanol and 0.5% Reagent DX before homogenization. The homogenate was centrifuged to remove any potential tissue residues and RNA from the clear supernatant was extracted using the Qiagen RNeasy Plus Mini Kit. RNA was quantified using NanoDrop (ThermoFisher) and RNA from 12 individuals was pooled to 3mg per pool (250ng per tissue sample). For library construction, the TruSeq Total RNA Library Preparation Protocol was used. Host ribosomal RNA (rRNA) was depleted using the Ribo-Zero-Gold Epidemiology Kit. Paired-end (100bp) sequencing of the RNA library was performed on the HiSeq 2500 platform (Illumina).