title: GSM8262140: Fish16; Salmo salar; RNA-Seq
library_construction_protocol: RNA was isolated with 900 µL QIAzol lysis reagent (Qiagen, Hilden, Germany, cat. No. 79306) and a 5 mm  stainless steel bead (Qiagen, cat. No. 69989). The tissues were lysed with a Qiagen TissueLyser bead mill  (Retsch, Haan, Germany, cat. no. 85220)  at 20 Hz for 2 min (twice). 180 µL chloroform was added before  vortexing. Tubes were centrifuged at 12 000 rpm (4 °C) The upper phase containing RNA and water was  transferred to new tubes, and 450 µL isopropanol was added. Tubes were shaken by hand and incubated  for 10 min at room temperature, before centrifugation for 15 min. Excess liquid was removed and 200 µL  70 % ethanol was added before centrifugation at 10 min. The supernatant was removed, and the samples  were centrifugated for 3 min. With the lids open, the pellets dried for 5-10 min. The RNA was then  dissolved in 40 μL RNAse-free water, followed by a 10 min incubation at 55 °C. 20 ng/ul. Poly A enrichment  and reverse transcription of cDNA.