Background: Hospitals have a large role in developing and maintaining public health. Hospital service lies in its human resources namely employees. Employee performance is influenced by work motivation and the existing organizational culture. Employee contributions are the main determining factor of organizational success. However, there are still employees who lose their motivation to work. Changes in the environment and intense competition make service quality reduce and this needs improvement. The purpose of this study was to analyze work motivation and cultural organization on the performance of employees RSUD dr. Achmad Diponegoro Method: The type of this research is quantitative research with cross-sectional approach. The research sample is as many as 137 employees taken by using purposive sampling techniques. The research instrument uses a questionnaire that has been tested for validity and reliability. The questionnaire consisted of 12 statements on motivation, 6 statements on organizational culture and 12 statements on performance. Data analysis using multiple linear regression. Results: This research obtains a work motivation hypothesis test with a significant level of 0,000 which indicates that there is an influence of work motivation on performance. The hypothesis test of organizational culture with a significance level of 0.083 shows that there is no influence between organizational culture on performance. The value of F count is 20,719 and significance is 0,000 show that there is a simultaneous influence between work motivation and organizational culture on employee performance. Conclusion: Work motivation influences employee performance, organizational culture does not influence employee performance, work motivation and organizational culture together have a positive and significant effect on employee performance.