title: GSM4586016: SAP_6h_3; Rattus norvegicus; RNA-Seq
library_construction_protocol: Total RNA of each lung tissue sample was extracted using TRIzol Reagent (Ambion) according to the manufacturer's instructions. To eliminate any DNA present, RQ1 DNase (Promega) was added to the extracted total RNA. The absorbance of the purified RNA was measured to determine the quality and concentration with a Smartspec plus (BioRad, Hercules, USA) at 260 nm/280 nm (A260/A280). Agarose gel electrophoresis (1.5%) was conducted to verify the integrity of the RNA For RNA-seq library preparation, 10 μg of purified total RNA of each sample was purified and concentrated with oligo (dT)-conjugated magnetic beads (Invitrogen). Polyadenylated mRNAs were used for the construction of a directional RNA-seq library. Ion fragmentation of purified mRNAs was conducted at 95ºC following end repair and 5' adaptor ligation. The fragmented mRNAs were reverse-transcribed with RT primer, known 3' adaptors, and random hexamers. Then, purification and amplification of the cDNAs were conducted. The 200–500 bps of amplified cDNAs were collected. The cDNAs for each sample were quantified and stored at -80ºC until sequencing.