The genetic variations in kappa-casein gene locus exon TV of three buffalo breeds, i.e. South Kanara, Surti, and Murrah, were investigated. PCR amplification of the kappa-casein gene using primers specific for Bos taurus yielded DNA products of a similar size. Upon restriction digestion with Hinf 1, two fragments having 266 bp and 84 bp were obtained for all the samples studied. Single strand conformation polymorphism (SSCP) and nucleotide sequence analysis also did not reveal genetic variation. BLAST alignment of 350 bp nucleotides of buffalo kappa-casein gene showed homogeneity with the kappa-casein gene of Syncerus caffer nanus. Analysis of the kappa-casein gene using PCR-restriction fragment length polymorphism and SSCP techniques including DNA sequencing data revealed a monomorphic pattern which is buffalo specific.