GSE112295
Active translatome profiling with RiboLace in MCF7 cells
We developed RiboLace, a novel method based on a new puromycin-containing molecule, for the isolation of active ribosomes and associate RNAs by means of an antibody-free and tag-free pull-down approach. RiboLace is fast, it works with very low input material and can be easily and rapidly used to obtain a transcriptome-wide snapshot of active translation from eukaryotic in vitro and in vivo systems. Keywords: translation, RiboLace, polysome profiling, RNA-seq, POL-seq, active translation Overall design: We performed RiboLace sequencing, polysomal RNA sequencing (POL-seq), total RNA sequencing (RNA-seq) and sequencing of the pull-down with mP-beads. Experiments were performed in duplicate on MCF7 cells in three conditions: control, serum starved (12h), serum starved and EGF treated (12h+4h).