Instrument: Illumina HiSeq 2500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: cDNA
Layout: SINGLE
Construction protocol: ribosome-protected fragments were isolated after snap-freezing of HeLa cells and Rnase I digestion using gel filtration columns. Gel purified fragments between 26 nt and 34 nt were isolated, rRNAs were removed using the RiboZero kit (Illumina). RNA-seq libraries were prepared from cells lysed similar as the ribosome profiling samples. totalRNAs was isolated from cleared lysates by addition of TriReagent as for the ribosome-protected fragments. Sequencing libraries from ribosome footprints were generated as previously described (Aeschimann et al., 2015). Total RNA was used for library generation with the TruSeq Stranded mRNA Library Prep Kit (Illumina). Libraries were sequenced on an Illumina HiSeq2500 generating 100 nt single-end reads.