The potential use of random amplified polymorphic DNA (RAPD) was evaluated as a source of development of alternative genetic markers for studying variation in buffalo (Bubalus bubalis) and other related species of the Artiodactyla family Bovidae, in order to ascertain genetic relationships and diversities. Fourteen arbitrary primers were used to amplify DNA fragments in four species such as Indian Zebu cattle (Bos indicus), buffalo (Bubalus bubalis), sheep (Ovis aries) and goat (Capra hircus). Clear and distinct RAPD patterns with a higher level of polymorphism was detected between species, while fewer polymorphisms were found within the species. Species were subsequently scored for presence or absence of RAPD fragments and Jaccard's similarity coefficients were calculated to quantify the genetic divergence among the species. Wagner parsimony analysis of the RAPD data for 542 markers resulted in one most parsimonious tree which revealed very low similarity among the four species analysed.