To characterize, identified and precisely diagnose liver flukes genetic variability among population has great importance. Several molecular techniques based on DNA have been applied to molecularly characterized Fasciola isolates, but there is no data about genetic analysis of Fasciola isolates from District Mardan Pakistan. In current study Fasciola isolates of sheep and goat collected from District Mardan of Pakistan were genetically analyzed by amplifying ITS1 gene of rDNA through PCR technique. Band pattern of all isolates showed a single band of ~470bp in length indicating that there is no variation in ITS1 region size between sheep and goat isolates. Sequencing analysis also confirmed nil variation.