title: RNA-seq of post-larval Spirinchus thaleichthys: whole fish (pooled sampled to assist in transcriptome assembly): Sample Longfin smelt_FCCL
design: "Four fish collected from the UC Davis Fish Conservation and Culture Laboratory that were from a different cohort (2013) as those used for the differential expression study were sequenced to aid in transcriptome assembly. Whole fish were homogenized using a TissueLyser (Qiagen) in Buffer RLT Plus (RNeasy Plus Mini Kit; Qiagen) and 350 ?l of the homogenate was used for the RNA extraction on a Qiacube (Qiagen) following manufacturer’s protocols. RNA quality was assessed using a Bioanalyzer (Agilent) and concentration was measured using a Qubit Fluorometer (Life Technologies). 1.5 ?g of total RNA from each fish was pooled and used to generate one Poly-A isolated, strand-specific cDNA library using a Directional PrepX RNA Library Prep kit (Wafergen Biosystems). This library for longfin smelt was pooled with the library for the wild fish and sequenced on an Illumina HiSeq 2500 over two lanes. Sequencing was conducted to produce 100 base pair paired-end reads. "