This benchmark document Playbook for Urban Biodiversity was produced by the Melbourne Centre for Cities at the University of Melbourne, as a key output from the Urban Nature: Urban Myths Symposium co-hosted with the City of Melbourne – 11 and 12 May 2023. This Playbook is intended to inform practice and policy on biodiversity in cities, with a particular focus on Melbourne and other Australian cities, using content presented by collaborating authors at the Symposium, supplemented by an extensive literature review. Published by Melbourne Centre for Cities, 2023.