This study aims to determine the relationship of independent variables: Organizational Culture,Transformational Leadership, and Work Motivation either individually or together with the dependent variable of Job Satisfaction. The selected sample is 249 people with criteria of lecturer of a private Islamic university in Jambi Province, Indonesia. This research uses Sequential Explanatory Design Mixed Method with quantitative method first then followed with the qualitative approach. This procedure is intended to produce more in-depth and comprehensive research. The result of this research concludes that: there is relationship of organizational culture with job satisfaction through regression equation Y = 13,126 + 0,880 X 1 with R2 = 76,8%, transformational leadership with job satisfaction through regression equation Y = 24,857 + 0,759 X 2 with R2 = 57,1 %, and work motivation with job satisfaction through regression equation Y = 28,384 + 0,747 X 3 with R2 = 55,6%. Also, there is positive relation of organisation culture, transformational leadership and work motivation together with job satisfaction through regression equation Y = 0,623 + 0,650 X1 + 0,149 X2 + 0,205 X3 with R2 = 83,2% indicating there are other factors that also related positive with an increase in job satisfaction of 16.8%.