RNA of Achirus lineatus:adult:gill waf exposed
To obtain a wide view of the differential transcriptome following 50% - WAF exposure, high-throughput RNA-Seq was performed using Illumina sequencing technology. First, total RNA was extracted by duplicate for both samples (~50 mg of liver and gill) from control and 50% WAF exposed fish, respectively. Total RNA extraction was performed using Direct-zol RNA MiniPrep kit (Zymo Research, U.S.A). The RNA quantity was assessed using NanoDrop 2000 (Thermo Fisher Scientific Inc., USA). The RNA integrity number (RIN) of each sample was measured using the Agilent Bioanalyzer 2100 system (Agilent Technologies, USA) equipped with an Agilent RNA 6000 Nano. All samples with values of RIN > 7.8 were selected for sequencing. Libraries construction and sequencing were performed in the Sequencing and Polymorphisms Identification Unit of the National Institute of Genomic Medicine (Mexico). cDNA libraries were constructed using the Illumina TruSeq Stranded mRNA sample prep kit (Illumina, Inc., SanDiego, CA). Products were quantified using the Agilent DNA high Sensitivity chip on the Agilent Bioanalyzer 2100 system. Libraries were quantified with the Qubit fluorometer using the Qubit ds DNA HS Assay (Invitrogen by Thermo Fisher Scientific, USA). The libraries were sequenced on an Illumina NextSeq500 platform, using the kit NextSeq 500/550 High Output v2 (300 cycles). Sequencing were done in a 2x150 bp paired end (PE) configuration
SUB7770043
gill_exposed_2