Introduction: The medical profession is a noble profession and it is a stressful job as well. Doctors experience extreme physical or mental tiredness due to their extended working hours, night shifts, increased workload, on-call duty, and less flexibility at work, which eventually significantly impacts their Work Life Balance (WLB) and Job Satisfaction (JS). Prevalence and relationship study about WLB and JS among medical officers has not been undertaken in the Sri Lankan context. This study focused on describing the prevalence of WLB and its relationship with JS among government medical officers in the Ampara district. Objectives: To describe the levels of WLB and JS, their association and the impact of WLB on JS among government medical officers in Ampara district. Methods: A descriptive cross sectional study was performed among 424 government medical officers working in hospitals using a stratified random sampling technique. A pre-tested and self-administered questionnaire was used and data were analyzed using SPSS. Pearson’s correlation coefficient, linear regression, and chi-square tests were deployed to examine the relationship between WLB and JS. Results/analysis: Out of 366 respondents, 66.1% and 17.8% of medical officers were having moderate and high levels of WLB respectively. Whereas 60.7% and 23.5% had moderate and high levels of JS respectively.  WLB showed a significant association with and impact on JS. Conclusions: This study provided a good understanding regarding the prevalence of the WLB and JS among government medical officers in Ampara district, Sri Lanka. A significant proportion of medical officers (66.1%) with a moderate level of WLB are at risk of developing low satisfaction in the future and significantly low prevalence of WLB and JS would affect the performance, well-being, patient care, and satisfaction if proper policies and programs are not incorporated. Good WLB and JS are important for medical officers to enhance the quality of life and quality of care.