Fig. 1: Schematic of Ribo-ITP.
From: Single-cell quantification of ribosome occupancy in early mouse development
a, Schematic of the generation of RPFs. Following RNase digestion, RPFs are isolated with the conventional or Ribo-ITP approach. b, Schematic of the conventional ribosome profiling protocol and the Ribo-ITP process for extraction of RPFs. In Ribo-ITP, marker oligonucleotides with a 5′ fluorophore (green circle) and 3′ ddC blocking modification (black circle), which encapsulate the size range of RPFs, are added to the digested cellular lysate. Lysate contents are loaded into the channel (t0), then an electrical current is applied to a selectively focus species of a specific electrophoretic mobility range, enabling nucleic acid extraction by ITP. Nucleic acids are extracted in a narrow ITP band and then size selected as they migrate through 5% (t1) and 10% (t2) polyacrylamide gels, respectively. At the end of the run, purified and size-selected RNAs are collected (t3).
