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Background: A number of factors (first experience, treating clinician and time to place dental-dam) may influence patients&rsquo, preferences regarding dental-dams. In general, patients accept placing it and that it must be used for teeth isolation during endodontic procures for the sake of patient safety and infection control. Objectives: The aim of this study is to investigate preferences and experiences of patients using dental-dam (DD) isolation during root canal treatment (RCT) and to explore influencing factors among the residents of Madinah Munnawara, Saudi Arabia. Methods: Following an ethical approval and a pilot study, a self-administrated questionnaire was distributed to 305 patients attending endodontic clinics at the Taibah University College of Dentistry (TUCOD) over six months. Patients voluntarily participated in the study after understanding the methodologies and signing a consent form. They were asked to fill out a questionnaire on their experiences and preferences in placing the DD during RCT. Data were analyzed using the Chi-square test at p = 0.05. Results: The response rate was 91%. There was no significant correlation between patients&rsquo, preferences and their race, age and gender (p &gt, 0.05). The majority of participants (74.3%) would prefer to use a DD in their next session (p &lt, 0.001). This preference negatively correlated with the time required to place a DD and the duration of the current visit (p &lt, 0.001). While most of those who would prefer to use a DD in their next visit were pleased with how it was placed in the current session (76.6%), most of those who would not do so (66.7%) were uncomfortable. Overall, the highest proportion of participants (40.2%) reported that prevention of instrument swallowing was the most important advantage of DD isolation (p &lt, 0.001). Conclusions: Overall, DD isolation for RCT is generally well accepted by patients regardless of their country of origin, gender, education and awareness of its advantages. Patients&rsquo, safety was the most attractive advantage for patients to the application of the DD. The time required to place the dental dam and first visit experience in placing the DD affect patients&rsquo, future preference.