The water buffalo holds tremendous potential in livestock sector, but we are defficient in genetic data of different Pakistani buffalo breeds, which needs to be established for their genetic identification. In this study, blood samples of unrelated true representative of two breeds (Nili- Ravi, Kundi) were collected from different government livestock farms.Mitochondrial Cytochrome b gene coupled with microsattelite markers are considered to be efficient tools for breed characterization & species identification.Extraction of DNA followed by polymerase chain reactions were performed for the partial amplification of mtCytb gene & nine microsatellite loci with specially designed primers on each DNA sample.Sequencing & genotyping were conducted, which helped us to find out SNPs to design different haplotypes for phylogenetic analysis & for allelic frequencies respectively. The results obtained in the current study must be beneficial in animal forensics for litigation & expedite the police investigation services in the region, which will also be useful for breed characterization & to know the evolutionary history of aforementioned buffalo breeds.
