Pakistan has vast population of dogs belonging to different breeds.Most of the dogs have no pedigree record which is a great threat to conservation of different breeds.No study on DNA fingerprinting of dogs has been conducted in Pakistan.DNA fingerprinting of dogs is necessary to overcome the problems like forensic cases, individual identity in case of fertilization by more than one male and ownership disputes.Microsatellite markers have been proved as an efficient tool for parentage testing and breed characterization of dogs. In this study, a panel of microsatellite markers was developed.Blood samples were taken from cephalic vein of two breeds of dogs(German shepherd and Labrador retriever). DNA was extracted and primers were optimized.Multiplex PCR was performed and genotyping was done for the PCR products on non denaturing polyacrylamide gel.These results were analyzed statistically.Almost all of the microsatellite markers showed significant variations in both breeds.Results of this study lead to development of a panel of microsatellite markers which can be used for parentage analysis and breed characterization of dog