GSM3259567: A549 cells transfected with GFP vector, replicate 2; Homo sapiens; RNA-Seq
RNA was extracted using the RNeasy Mini Kit (QIAGEN) according to the manufacturer's instructions, adding 600 μls RLT Buffer and 6 μls of B-mercaptoethanol to each sample. One ug of DNase-treated RNA was depleted of rRNAs using the Ribo-Zero Gold rRNA  Removal Kit (Human/Mouse/Rat) according to the manufacturer's instructions and purified  post-depletion with 1.6X volume AMPure XP beads. Barcoded directional RNA-Seq libraries  were then prepared using the TruSeq Directional Library Prep kit (Illumina), per kit  instructions. PCR products were purified with 1.8X volume AMPure XP beads and fragments  of 300-500 bp were size-selected using BluePippin 2% M1 gels (Sage Scientific). Afterwards,  libraries were sequenced on the Illumina HiSeq 2500 platform in a 100 bp single-end read run  Format.