This study aimed to compare the efficacy of robot-assisted and manual cannula insertion in simulated big-bubble deep anterior lamellar keratoplasty (DALK). Novice surgeons with no prior experience in performing DALK were trained to perform the procedure using manual or robot-assisted techniques. The results showed that both methods could generate an airtight tunnel in the porcine cornea, and result in successful generation of a deep stromal demarcation plane representing sufficient depth reached for big-bubble generation in most cases. However, the combination of intraoperative OCT and robotic assistance received a significant increase in the depth of achieved detachment in non-perforated cases, comprising a mean of 89% as opposed to 85% of the cornea in manual trials. This research suggests that robot-assisted DALK may offer certain advantages over manual techniques, particularly when used in conjunction with intraoperative OCT.