Separation of chromatin containing bromodeoxyuridine in one or both strands of the DNA.

A method has been devised for the separation of chromatin containing 5-bromodeoxyuridine (BrUdRib) in one strand (HL) of the DNA from that with BrUdRib in both strands (HH). Ultraviolet light breaks chromatin containing HH DNA into smaller fragments than chromatin containing HL DNA and the two species can be partially resolved on neutral sucrose gradients. Unfiltered ultraviolet light is not suitable since it causes considerable alteration in the electrophoretic pattern of chromatin-associated proteins. Irradiation with 313-nm light causes much less damage to the associated proteins. The ability to separate, isolate, and examine chromatin containing HL and HH DNA makes studies on the distribution of chromatin-associated proteins possible.