Chronic wounds develop as a result of defective regulation in one or more of the complex cellular and molecular processes involved in proper healing. They impact 6.5M people and cost $25B/year in the US alone. Although a significant effort has been invested in understanding how chronic wounds develop in humans, the work has met with little success. This is primarily because we cannot experiment in humans and because of the lack of proper animal models to conduct studies on the processes that lead to the onset of chronicity. We have recently developed a novel mouse model for diabetic chronic wounds that have many of the characteristics of human chronic wounds. Using db/db mice, we can generate chronic wounds 100% of the time by inducing high levels of oxidative stress (OS) in the wound tissue immediately after wounding, using a one-time treatment with inhibitors specific to the antioxidant enzymes catalase and GPx. These wounds have high levels of OS, develop biofilm naturally, become fully chronic within 20 days after treatment and sometimes remain open more than 100 days if the mouse survives that long. This novel model has many features of diabetic chronic wounds in humans and therefore it will contribute significantly to advancing fundamental understanding of how wounds become chronic. This is a major breakthrough because chronic wounds in humans cause significant pain and distress to patients, many of whom result in amputated. Moreover, these wounds are very expensive and time-consuming to treat and lead to significant loss of personal income to patients. Advancements in this field of study through the use of our chronic wound model can significantly improve health care for millions who suffer under this debilitating condition. The protocol we will submit here describes in great detail how we perform the procedure to cause acute wounds to become chronic, something that was not possible when we published the scientific paper presenting this model (Dhall et al., Dec. 2014).