GSM5543656: rainbow trout gill cell line RTgill-W1 (ATCC®CRL-2523™) 100 mg/L LPS-treated Replicate2; Oncorhynchus mykiss; RNA-Seq
Cells were washed with PBS and the plates stored at -80°C upon completion of treatment protocol. Upon thawing, total RNA was extracted from each replicate well using RNeasy Plus Mini kits (QIAGEN), following the manufacturer's protocol. Only samples with an RNA Integrity Number (RIN) > 8 were used for dsDNA library preparation. Libraries were prepared using the TruSeq® Stranded mRNA Library Prep kit (Illumina Inc, San Diego CA, USA) and were subsequently run for paired-end sequencing (75 bp x 2, 150 cycles) using the NextSeq 500/550 High Output Kit v2.5 (Illumina Inc) on an Illumina NextSeq 500 System (Illumina Inc). Sequencing was conducted on four control replicates and three 100 mg L-1 LPS treatment group replicates. Only samples with an RNA Integrity Number (RIN) > 8 were used for dsDNA library preparation. Libraries were prepared using the TruSeq® Stranded mRNA Library Prep kit (Illumina Inc, San Diego CA, USA). The quality of dsDNA libraries were assessed using Agilent high sensitivity DNA analysis kit in a 2100 Bioanalyzer instrument (Agilent Technologies, Waldbronn, Germany). Libraries were quantified in a Qubit 4 fluorometer (Thermo Fisher Scientific).