Backgrounds and study aims: Rescue techniques applied in case of cannulation failure are limited in number and have severe limitations. We recently described partial ampullary endoscopic mucosal resection (PA-EMR) as a novel technique for difficult biliary cannulation. We aimed to demonstrate the efficacy and safety of PA-EMR for patients with difficult biliary cannulation in a larger cohort Patients and methods: We conducted a prospective cohort study at our tertiary-care referral hospital. We recorded demographic factors, procedure-related characteristics, and adverse event rates in patients who received PA-EMR in case of difficult biliary cannulation. Results: Between June 2021 and June 2022, we performed 1073 consecutive ERCP procedures on 962 patients; 40 patients (12 male, 28 female, with a mean age of 68.1± 4 years) had difficult biliary cannulation and had PA-EMR. Technical success was 100%, and none of the patients required a second session. None of the patients had an adverse event, and we encountered no 30-day mortality. The most typical indication was bile duct stones with 60% (n=24), followed by periampullary tumors with 15% (n=6). The total procedure time was 923 (min-max, 392-1224) seconds. Conclusions: PA-EMR is an effective and safe rescue technique in patients with difficult biliary cannulation.