Ecosystem fragmentation by human intervention breaks down the biosphere habitat. Wildlife corridors connect biosphere habitats to maintain ecosystem continuity and provide animals with connecting routes. In Korea, there are 17 existing wildlife corridors on highways (as of December, 2006. Korea Freeway Corporation). There are 24 highway routes, 2,923km of highway(as of December, 2004. Korea Freeway Corporation). However, wildlife corridors are not enough and roadkill increases every year, so we need to construct additional wildlife corridors on highways. This study proposes a new approach to the location of wildlife corridors on highways, using a comprehensive analysis method for main location elements, and applies it to a study area. First, it examines traditional approaches to location of wildlife corridors through literature review and field study to analyze the present conditions of existing wildlife corridors. Then, it developes a comprehensive analysis method for the location of wildlife corridors. (1) Field investigation : investigate planting, water bodies and so on. (2) Roadkill analysis : roadkill counts, locations, time, and so on. (3) Monitoring : animal traces were surveyed and sensor cameras were installed to determine target species. (4) Simulation for animal movement : most probable wildlife dispersal was simulated by a computer software. (5) A new comprehensive approach overlays all analysis on a map and determines the location of proposed new wildlife corridors. In conclusion, it proposes an over-bridge type wildlife corridor in Dalnaenae Hill (413-414km from Busan) and an underpass type near the entrance to Gwanhyun temple way (415-416km from Busan). This new approach based on roadkill data, computer simulation of wildlife dispersal, monitoring of animals, and site analysis, may contribute to better location of wildlife corridors on highways.