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UV Laser-Induced Protein-DNA Crosslinking

  • Stefan I. Dimitrov
  • Tom Moss
Part of the Methods in Molecular Biology™ book series (MIMB, volume 30)

Abstract

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, 2, 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).

Keywords

Potassium Thiocyanate Prehybridization Buffer Antibody Buffer Chemical Crosslinking Agent Photochemical Crosslinking 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc. 1994

Authors and Affiliations

  • Stefan I. Dimitrov
    • 1
  • Tom Moss
    • 1
  1. 1.Centre de Recherche en Cancérologie deI'Université LavalQuébecCanada

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