Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: X. maculatus at varies at varies age were sacrificed for RNA isolation. Total RNA was isolated using TriReagent (Sigma Inc., St. Louis, MO, USA). Tissues from each individual fish was homogenized in Tri-reagent followed by addition of 200 μl chloroform and the samples vigorously shaken and subjected to centrifugation at 12,000 g for 15min at 4 °c. Total RNA was further purified using RNeasy mini RNA isolation kit (Qiagen, Valencia, CA, USA). Residual DNA was eliminated by performing on column DNase digestion at 37°C for 30 min. RNA integrity was assessed by gel electrophoresis (2% agarose in TAE running buffer) and total RNA concentration was determined using a spectrophotometer (NanoDrop Technologies, Wilmington, DE, USA). RNA quality was verified on an Agilent Bioanalyzer (Agilent Technologies, Santa Clara, CA) to confirm that RIN scores were above 8 prior to sequencing. RNA libraries were prepared for sequencing using standard Illumina protocols. Libraries were constructed using the Illumina TrueSeq library preparation system that employs a polyA selection