Affinity chromatography with biotinylated RNAs

Abstract

Small, reversibly biotinylated RNAs as described here are versatile ligands for affinity chromatography of RNA-binding components. These RNAs can be attached to a solid support by binding to avidin and used as ligands, or they may be hybridized to another RNA which acts as the ligand. The incorporation of a disulfide bond in the linker arm connecting biotin to the RNA makes it possible to dissociate the RNA from avidin under mild conditions. Our results regarding the binding and elution of the biotinylated RNA may be applied to other, reversibly biotinylated molecules.