title: RNAseq of mus musculus: lung tissue: 24hr post exposure: rep3: Aerosol exposure
design: Each experimental time point for the chosen studies contained four to five replicates. Dissected lung tissue was stored in RNAlater and frozen at -80 until RNA isolation. Each dissected lung was moved separately to a Bead Beater tube containing 600ul of buffer RLT plus from Qiagen RNAeasy Mini Kit (ThermoFisher). Tissues were homogenized for one minute in a mini-beadbeater-24 and subsequently centrifuged for three minutes to reduce foaming. After spinning, RNA was extracted using the RNAeasy Mini Kit following the manufactures protocol. RNA was stored at -80 until further processing. Total RNA was quality checked and quantified using a BioAnalyzer 2100 (Agilent) and determined to be of good quality. A total of 45 (21 aerosol treated and 24 intranasally treated) sequencing libraries were constructed using TruSeq RNA sample prep kit v2 (Illumina). For each treatment (aerosol/intranasal) each library was indexed and sequenced on a single lane on the HiSeq2500 platform (Illumina), producing 10-30 million 150bp single-end reads for each replicate.