GSM5655864: TRA_72h_3; Trachinotus blochii; RNA-Seq
The total RNA was harvested using Trizol-LS reagent. Total RNA was extracted from liver and brain using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's directions. RNA samples were digested using DNase I to remove potential genomic DNA and stored at -80℃. RNA was quantified and quality was assessed by NanoDrop Spectrophotometer 2000c (NanoDrop Technologies, Wilmington, DE, USA) and 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA), and all samples were standardised to 500 ng/μl. Three samples from per group at each time point were used for RNA library construction which OD ratios of 260/280 and 260/230 greater than 1.8 and RNA Integrity Number (RIN) greater than 8. RNA-seq library preparation and sequencing were carried out by Annoroad Gene Technology Co., Ltd (Beijing, China), forty five sequencing libraries were constructed using a TruSeq™ RNA Sample Preparation Kit (Illumina) according to the manufacturer's directions. The next step was KAPA quantifcation and dilution, after which the library was sequenced on an Illumina HiSeq.2500 with 125-bp paired-end reads. After removing adaptor sequences, ambiguous N nucleotides (those an N ratio <5%) and low quality sequences (reads with <50% bases of quality value), the remaining clean reads were assembled using Trinity sofware