Name: Ribosome profiling (RiboSeq) of ribosome footprints from LoVo cells at time 2h post-infection), replicate 2
Instrument: NextSeq 500
Strategy: RNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: SINGLE
Construction protocol: The methodology employed for generating ribosome footprint libraries was based on the original protocols of Ingolia and colleagues (Ingolia et al., 2012 doi:10.1038/nprot.2012.086), except that library amplicons were constructed using a strategy inspired from iCLIP experiments (Huppertz et al., 2014 doi: 10.1016/j.ymeth.2013.10.011, Haberman et al., 2017 doi:10.1186/s13059-016-1130-x) and adapted to Illumina NextSeq500. Mainly, in this protocol, the reverse transcription primer contains not only an external identifier barcode for library demultiplexing, but also a unique molecular identifier allowing the identification and removal of PCR duplicates.