[Objective] The aim was to improve the biogas production rate,and improve the use ratio of rice straws to the furthest.[Method] Single-factor method was adopted to analyze the influence of inoculated amount,total solid content and carbon-nitrogen ratio on biogas production in biogas station of Nanjing University of Technology.[Result] It was found that the most optimum condition was:inoculated amount of 34%,total solid content of 6%,and carbon-nitrogen ratio of 25:1.According to the experimental verification,the total biogas yield in experimental group increased by 27.6% over the control group.[Conclusion] The research provided the reference of the further industrialization production of biogas using straw.