Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: PAIRED
Construction protocol: Total RNA was isolated from the human respiratory sample with the QIAGEN RNeasy Micro Kit Double-stranded cDNA was synthesized and amplified from 200-500 picograms of input RNA with the NuGEN Ovation RNA-seq V2 kit and purified using the QIAquick PCR Purification Kit. cDNA was sheared to 250 bp using the S220 Focused-ultrasonicator and purified using Agencourt RNAClean XP beads. Sequencing libraries were prepared from 100 ng of purified, sheared cDNA with the NuGen Ovation Ultralow System V2 1-16. The libraries were sequenced on the Illumina HiSeq 2500 instrument with TruSeq SBS V3 chemistry according to the manufacturer's instructions.