On-site construction activities can result in significant mobility and safety impacts to road users. The presence of work zone can also result in inconvenience to local business and community, noise and environmental impacts. Minimizing the adverse impacts of work zones has become a higher priority especially since the inception of the FHWA Rule on Work Zone Safety and Mobility (23 CFR 630 Subpart J). Work zone road user costs (WZ RUC) provide the economic basis for quantifying these adverse impacts which can then be used for effective decision-making to improve work zone mobility and safety. This report provides practitioners with information on WZ RUC analysis concepts and their applications using case studies drawn from real world projects. WZ RUC primarily refers to monetized components of mobility and safety impacts; increasingly, non-monetary and qualitative components, such as environmental, business, and societal impacts, are being utilized. In this report, each of the monetary components is explored and the computations of these components are illustrated using examples. It presents step-by-step procedures to derive unit costs for monetary components. It lists the cost sources for each cost component as well as the ways to update those using economic indices. It also explores input requirements and various tools available for use in WZ RUC analysis. This report presents the application of both monetary and qualitative components of WZ RUC in MOT alternative analysis using a decision analysis framework. Another key application of WZ RUC is in selecting appropriate project delivery/contracting strategies to minimize WZ RUCs and related impacts through early project completion. Approaches for determining an appropriate level of incentives and disincentives are also discussed. Three “real-world” case studies from the Highways for LIFE program are presented to demonstrate the applications of WZ RUCs in selecting the preferred Maintenance of Traffic (MOT) alternatives and project delivery/contracting strategies.