GSM6260410: T3008.Control.AEC.Alone; Homo sapiens; RNA-Seq
Following infection of AECs with HRV16 for 48 hours, RNA was extracted from the AECs and LAD2 MCs using a RNAqueous™-Micro Total RNA Isolation Kit (ThermoFisher/Life Technologies). Using 1ug total RNA with a RNA integrity number (RIN) ≥8, polyadenylated RNA (primarily mRNA) was selected and purified using oligo-dT conjugated magnetic beads. cDNA libraries were then constructed for each sample using the TruSeq Stranded mRNA Sample Prep Kit (Illumina, #RS-122-2103). Final cDNA libraries were analyzed for size distribution using an Agilent Bioanalyzer (DNA 1000 kit, Agilent # 5067-1504), quantitated by qPCR (KAPA Library Quant Kit, KAPA Biosystems # KK4824), then normalized to 2 nM in preparation for sequencing. Libraries were pooled and quantified using a Qubit® Fluorometer (Life Technologies). Sequencing of pooled libraries was carried out on a HiSeq 2500 sequencer (Illumina) with paired-end 53-base reads and a target depth of 5 million reads per sample.