Laser interference lithography is an effective approach for grating fabrication. As a key parameter of the grating profile, the duty cycle determines the diffraction characteristics and is associated with the irradiance of the exposure beam. In this study, we developed a fabrication technique amplitude-splitting flat-top beam interference lithography to improve duty cycle uniformity. The relationship between the duty cycle uniformity and irradiance of the exposure beam is analyzed, and the results indicate that when the beam irradiance nonuniformity is less than 20%, the grating duty cycle nonuniformity is maintained below ±2%. Moreover, an experimental amplitude-splitting flat-top beam interference lithography system is developed to realize an incident beam irradiance nonuniformity of 21%. The full-aperture duty cycle nonuniformity of the fabricated grating is less than ±3%. Amplitude-splitting flat-top beam interference lithography improves duty cycle uniformity, greatly reduces energy loss compared to conventional apodization, and is more suitable for manufacturing highly uniform gratings over large areas.