RNA-seq of Atlantic salmon: AGD-infected gill
A standard TRI Reagent RNA extraction  protocol was followed. Briefly, approximately 50 mg of tissue was homogenized  in 1 ml of TRI Reagent (Sigma, St. Louis, MO) by shaking using 1.4 mm silica  beads, then 100 l of 1-bromo-3-chloropropane (BCP) was added for phase  separation. This was followed by precipitation with 500 l of isopropanol and  subsequent washes with 65-75 % ethanol. The RNA was then resuspended in  RNAse-free water and treated with Turbo DNAse (Ambion). Samples were cleaned  up using Qiagen RNeasy Mini kit columns and their integrity was checked on  Agilent 2200 Bioanalyzer (Agilent Technologies, USA). Thereafter, the  Illumina Truseq mRNA stranded RNA-Seq Library Prep Kit protocol was followed  directly. Libraries were checked for quality and quantied using the  Bioanalyzer 2100 (Agilent), before being sequenced on three lanes of the  Illumina Hiseq 4000 instrument using 75 base paired-end sequencing at  Edinburgh Genomics, UK.
SUB5912490
513G