RNA-seq of Senegalese sole testis: comparison between fish fed a control and an algae-supplemented diet
RNA-seq of Senegalese sole testis: comparison between fish fed a control and an algae-supplemented diet
By the end of the experiment, 3 male fish per tank (2 tanks per group) were randomly sacrificed with a lethal dose of anesthesia (1000 ppm phenoxyethanol) and testis were immediately dissected and frozen in liquid nitrogen. Testis samples were stored at -80 ºC for RNA extraction and further RNA-Seq quantification. The gonads collected were homogenized in 1 mL of TRI Reagent® (Sigma-Aldrich, Switzerland) using a tissue-lyzer Star-Beater (VWR). Total RNA was extracted following the instructions of the manufacturer and the obtained pellet was resuspended in RNase-free distilled water (Sigma-Aldrich, Switzerland). Total RNA concentrations were measured with a Nanodrop One-C spectrophotometer (Thermo Fisher Scientific, Washington, USA) and adjusted to 1 µg/µL. Total RNA quality and integrity were checked using a 2200 TapeStation (Agilent Technologies, Santa Clara, CA, USA), and only samples with an RNA integrity number (RIN) > 8 were considered for sequencing. A total of 12 libraries were prepared using a TruSeq™ RNA Library Preparation Kit v2 of Illumina (Illumina, USA).