Abstract Sex identification was essential for the management, human-assisted breeding, prevention and cure of genetic disease and evolutionary studies of birds, especially for endangered birds. Red-Crowned Crane is a first class protected animal in China and a kind of monomorphic bird, so it’s difficult to identify the sex with morphological method, this situation challenged the couple and human-assisted breeding of Red-Crowned Crane. Genome DNA was extracted from feathers by using nondestructive sampling method and three combinations of primers were selected to amplify the EE0.6 sequence of Red-Crowned Crane specifically. The results showed that male birds displayed only one distinct amplification band, whereas two bands were amplified for female ones. With this molecular method in the article, the sex of Red-Crowned Crane could be identified successfully and accurately and then solved the key problem during the human-assisted breeding of Red-Crowned Crane. This method also gave a guide and reference for the accurate sex identification of other endangered birds.
