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UV Laser-Induced Protein-DNA Crosslinking

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Photochemical crosslinking is a powerful method for studying all types of protein-nucleic acids interactions. In particular, UV-induced crosslinking has been successfully applied to the study of protein-DNA interactions (1 ). Ultraviolet (UV) light is a zero-length crosslinking agent. It is therefore not subject to the steric problems that can be associated with chemical crosslinking agents and provides strong evidence for close protein-DNA interactions. However, to achieve an acceptable degree of crosslinking with conventional UV light sources, exposure times ranging from minutes to several hours have had to be used (1 3 ). Such prolonged irradiation allows for redistribution of proteins and the artifactual crosslinking of UV-damaged molecules, and it also precludes kinetic studies. The use of UV lasers overcomes these difficulties, since crosslinking is achieved after only nano- or picosecond exposures (4 5 ).
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