The Southern flying squirrel (Glaucomys volans) is a species designated at risk in Canada where its range is restricted to parts of Ontario, Quebec and Nova Scotia. Before this study, its distribution in Nova Scotia was poorly documented, with only seven site records. Based on live-trapping and intact and partial specimens provided by the public, we present data for 28 additional locations; these combined with historic records delineate a disjunct range that is more extensive than previously believed, but limited to southwest Nova Scotia. To identify specimens that were not fully intact, simple morphological and molecular techniques were employed. The latter, which consisted of PCR amplification and then restriction enzyme digestion of the cytochrome-b gene, allowed reliable species identification of tree squirrels from Nova Scotia by use of partial specimens.