Instrument: Illumina HiSeq 2000
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: 4 volumes of Trizol per volume of animals were added. The tubes were immediately frozen in liquid nitrogen, thawed and briefly vortexed; this step was repeated and then followed by a 5-minute incubation at room temperature. To each tube 0.2 ml of chloroform per 1 ml of Trizol was added, mixed gently, for 15 seconds, incubated at room temperature for 2-3 minutes, and then centrifuged at 12,000g for 15 minutes at 4ºC. The colorless upper phase was transferred to an RNase free tube. RNA was then purified using an Ambion PureLink RNA Mini Kit (Cat#12183018A). Following RNA purification, the RNA was treated with DNase I (Life Technologies Cat# 12185010), quantified using a NanoDrop Spectrophotometer and stored at -80ºC. RNA libraries were prepared for sequencing using standard Illumina protocols