To investigate the role of robotic single-site (RSS) ovarian cystectomy in fertility preservation, which was compared with single-port laparoscopic (SPL) surgery based on AMH changes. We retrospectively analyzed medical records of total 156 patients who underwent SPL (n = 72) or RSS (n = 84) surgery with the da Vinci Si or Xi system. The pre/post-operative AMH levels and total diameter of ovarian cysts were measured. In addition to the surgical method, AMH changes were compared according to the laterality, multiplicity, and pathology of ovarian cysts. A comparison of the characteristics of the SPL group and RSS group, revealed that there were no significant differences in the average age, the diameter of the ovarian cyst, and the number of locule. There were also no statistical differences between the pre-operative and post-operative AMH levels and the average surgical time including the docking time in robotic surgery. A comparison based on the surgical methods, revealed that the decrease in post-operative AMH was lower in the RSS group (24.2 ± 35.9%) than in the SPL group (34.9 ± 29.1%) significantly (p = 0.044). In patients with endometriosis, the decrease in AMH was greater, than that in patients without endometriosis. A longer operation time, larger ovarian cysts and multi-locular cysts were associated with lower AMH level in both the SPL and RSS groups (Pearson correlation coefficient: – 0.320, p = 0.0001, – 0.218, p = 0.007, – 0.236, p = 0.003, respectively). RSS ovarian cystectomy could be a promising new therapeutic option for fertility preservation in complex cases to avoid an additional side port.© 2023. The Author(s).