The Powerful Owl is an elusive species inhabiting the forests of mainland eastern Australia. Obtaining crucial information on aspects of their breeding behaviour and dispersal has proven extremely difficult, even though other aspects of their ecology are well studied. Here we use molecular methods to investigate the breeding behaviour and dispersal of the Powerful Owl in two different habitats: highly fragmented forest along the urban fringe and continuous forest. DNA profiles of Powerful Owls were obtained predominately from shed feathers collected opportunistically between 1995 and 2006. Seven breeding pairs of Powerful Owls were identified, from which shed feathers were collected during 2003, 2004 and 2005. By comparing DNA profiles, one pair of Owls was found to have occupied the same breeding site for 10 years (1995a??2005). The dispersal or movements of five offspring from this pair was also determined to be either of two scenarios: (1) the juvenile moves from the natal territory; however, isna??t breeding; and (2) the juvenile is recovered as part of a breeding pair. Two pairs of Owls breeding in the urban fringe habitat were closely related, but no incidences of extra-pair fertilisation were detected among pairs in either habitat. This study provides new information about the breeding behaviour and dispersal of the Powerful Owl, and shows the potential of using genetic data sourced from shed feathers for studying cryptic, rare or elusive species.