Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: Total RNA was extracted from 20 rat AEC-I samples and DNase treated before library preparation for next generation sequencing. We pooled samples from 4 technical replicates of monolayers prepared from the same animal to provide sufficient genomic material for RNA extraction. 500ng of total RNA was used to generate libraries with Lexogen's QuantSeq 3'mRNA-Seq library prep kit for Illumina, FWD (Cat #015.24). The QuantSeq kit generates one fragment per transcript and sequences obtained are close to the 3'end of polyadenylated RNA. The quality of purified libraries was checked by Agilent Bioanalyzer using High Sensitivity DNA kit and quantified by KAPA library Quantification kit_Illumina (Cat # KK4835). Finally, equimolar concentrations of all libraries were pooled and subsequently next generation sequencing was performed on the NextSeq500 instrument (75 bp single read) using NextSeq(tm) 500 Mid Output Kit (cat#FC-404-2001).