GSM7910690: liver/anterior intestine/gill, Day 14, 20 ppt, rep b; Trachinotus carolinus; RNA-Seq
In order to ensure sufficient RNA yields for transcriptomic analysis, the liver, anterior intestine, and gill lamellae (removed from three gill arches on the left side of the fish) were dissected and pooled in 1 mL cryovials, then flash frozen at -80oC. All samples were stored at -80oC until RNA extraction. RNA extractions were conducted on pooled gill lamellae, immune organs, and liver samples using Qiagen RNeasy kits (Qiagen, Hilden, Germany) according to manufacturer protocols. RNA quality and concentrations were determined using a Qubit Fluorometer 3 (Thermo Fisher Scientific, Waltham, MA) and a ThermoFisher NanoDrop Spectrophotometer 2000 (Thermo Fisher Scientific Inc., Waltham, MA). Extracted RNA for transcriptomics was flash frozen and shipped overnight on dry ice to Novogene (Sacramento, CA) for high throughput RNA sequencing. Total RNA was used to create 21 poly-A enriched libraries that were then sequenced on an Illumina NovaSeq to generate 2 x 150 bp paired-ends sequencing reads.