We report here on our technique and outcomes of the first two cases of robot-assisted laparoscopic nephroureterectomy with a bladder cuff excision (RLNU). RLNU was performed on two female patients who both had a muscle invasive lower ureter tumor. For the first step, nephroureterectomy was performed in the lateral flank position. For the second step, bladder cuff excision and bladder repair were performed in a steep Trendelenburg position. The specimen was extracted through a 6 cm sized incision in the umbilical trocar site. Both procedures were successfully completed with using the robot without conversion to open surgery. The total operative time, including the lymphadenectomy and the robot docking times, was 320 and 241 minutes, respectively, for the 2 patients. The estimated blood loss was 40 and 200 ml, respectively. The pathological examinations showed stage T3 and T2 invasive transitional cell carcinoma of the ureter. The patients' postoperative recoveries were uneventful and the bladder cuff was free of tumor. RLNU may have potential advantages over open and laparosopic surgery due to its minimal invasiveness. This approach can be an alternative to open surgery or laparoscopic technique.