Due to the potential risk of extinction of plant and animal species in Iran and Isfahan, ex-situ conservation strategies such as establishment of gene banks should be considered. It is therefore necessary to establish a gene bank of wildlife biological samples in Isfahan Province. The current project aimed to sample wildlife species inhabiting protected areas or other habitats within the province including mammals, birds and fish. Base on the species, appropriate sampling techniques such as trapping, use of fish net, etc were used. In the current project we collected 123 biological samples from wildlife species of Isfahan including 17 samples of 13 mammalian species, 70 samples of 35 bird species and 36 samples of 3 native fish. Common and scientific names of species, gender and sampling locations were recorded and biological samples were prepared for each species. Number of samples for each species varied from 1 to 3, though for fish species more than 3 samples were collected. A distribution map of sampling localities in the province was produced. Due to the seasonal variability of species presence and abundance, it is recommended to collect samples in different seasons to increase the number of samples