title: PTRE-seq in HeLa
design: Next generation sequencing library preparation was performed as previously described (Kwasnieski et al. 2012; White et al. 2013; Kwasnieski et al. 2014). RNA was isolated from cells using RNeasy Kit (Qiagen) or from polysome fractions with Trizol (Amresco).  RNA was treated with Turbo DNase (Life Technologies) and reverse transcription was performed using Superscript IV reverse transcriptase (Life Technologies). The 3  UTR of the luciferase gene containing barcode sequences was amplified (Phusion high fidelity master mix, New England Biolabs) from cellular or polysome associated RNA (cDNA) or plasmid DNA prior to next-generation sequencing. Each replicate experiment was sequenced as a pool on four lanes on an Illumina NextSeq sequencer. Illumina Single End