RNA-protein interactions were characterized using the iCLIP protcol (cross-linking and immunoprecipitation). RNA was UV-crosslinked to protein, isolated by immunoprecipitation using msl-2 antibody, and characterized by deep sequencing analysis.
Source was cell extract of clone8 cell line; bait produced from endogenous gene; prey produced from endogenous gene.
RNA bait was tagged with MS2 stem loops, and immobilized on amylose beads using a fusion of MS2-binding protein and maltose binding protein (MBP).
Interaction in vitro; bait produced by in vitro transcription; prey produced as a recombinant fusion protein in baculovirus-infected Sf21 cell system.