We isolated and characterized the first chromosome-specific satellite DNA (HC2sat) of Chinese hamster. This novel satellite was localized to the pericentric region of hamster chromosome 2. The 2.8 kb long repeat unit of HC2sat was identified and two such units were sequenced. Extended short range periodicity could not be revealed in repeat units. These elements are amongst the largest satellite repeat units reported from mammals to date. HC2sat is a major constituent of the pericentric region of CHO chromosome 2 representing a 7-14 Mb long DNA segment. Studies of long range organization of HC2sat indicated that the formation of the satellite array might occur in different phases and involved different amplification mechanisms.