Time-Resolved Hydroxyl Radical Footprinting of RNA with X-Rays

摘要

RNA footprinting by hydroxyl radical cleavage provides ‘snapshots’ of RNA tertiary structure or protein interactions that bury the RNA backbone. Generation of hydroxyl radicals with a high-flux synchrotron X-ray beam provides analysis on a short timescale (5 ms to 100 ms), which enables the structures of folding intermediates or other transient conformational states to be determined in biochemical solutions or in cells. This unit provides protocols for using synchrotron beamlines for hydroxyl radical footprinting.