Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Murine lungs were harvested immediately after the time of sacrifice, sliced into small pieces, homogenized using BeadBlaster Microtube Homogenizer (90sec, 4000rpm) in the presence of QIAzol. RNA isolation was then performed according to miRNeasy kit protocol (Qiagen). mRNA quality was checked using the Agilent 2100 Bioanalyzer according to the manufacturer's instructions. All RNA Integrity Numbers (RINs) were higher than 8. cDNA libraries were prepared using 2µg of isolated RNA according to the manufacturer's instructions of the SENSE mRNA-Seq Library Prep Kit V2 for Illumina (Lexogen). DNA size and quality were checked using the Agilent 2100 Bioanalyzer, according to the manufacturer's instructions. Libraries were quantified by using the Qubit DNA HS Assay kit (Invitrogen). The amplified libraries, each sample having its own index primer, were pooled at a total concentration of 2 nM and sequenced using the Illumina HiSeq platform (Technion Genome Center, Israel).