Background: Cost estimation is a central feature of health economic analyses. The aim of this study was to use a micro-costing approach of healthcare perspective to estimate the costs associated with conducting mass vaccination in order to be used for future campaigns in Jordan. Method: Micro-costing is a detailed method for evaluating the costs of a vaccination campaign. It begins by defining the campaign’s scope and objectives, followed by identifying all resources used—such as personnel, vaccines, and facilities. These are categorized into direct (material and labor) and indirect (overhead) costs. Resource use is quantified, costs are assigned accordingly, and total and per-unit (per dose) costs are calculated. Result: Costs of mass vaccination campaigns of inactivated was estimated to be 35,985,974.97 JOD. Specifically, the cost of overhead resources was 4,942,070.42 JOD. The staffing costs were 4,782,031.83 JOD while material costs were 26,269,026.24 JOD. Conclusion: Mass vaccination campaigns need good planning in order to use resources efficiently and effectively.