Procedures are described for the dissociation of histones H1 and H5 from chicken reticulocyte chromatin without disruption of the native core histone-DNA complex. The comparative properties of native and depleted chromatin with respect to sedimentation, thermal denaturation, and sensitivity to nuclease digestion have been studied. The changes in these properties resulting from removal of the linker histones are fully reversed when histone H5 is added back to the depleted chromatin.