RNA-Seq of Perca fluviatilis: spleen
Total RNA was extracted from the homogenized frozen tissues (whole eye, gill, spleen, liver and olfactory rosette; ~ 15-20 mg each) using NucleoSpin RNA extraction kit (MACHEREY-NAGEL, Duren, Germany). The quality of the total RNA sample was evaluated using TapeStation 2200 (Agilent) electrophoresis and sample concentration was measured with Nanodrop ND-2000 (Thermo Scientific). The preparation of RNA libraries and sequencing was performed at the Novogene Cambridge Science Park (Cambridge, United Kingdom). Libraries were prepared from total RNA by polyA capture with magnetic beads, followed by a reverse transcription of cDNA and second strand cDNA synthesis according to Novogene NGS RNA Library Prep Set (PT042) instructions (Novogene). After fragmentation and end repair, poly-A tail attachment, adapter ligation, size selection, amplification, and purification, the libraries were sequenced (2 x 150 bp design) using NovaSeq 6000 (Illumina) instrument.