Background: This survey study aimed at investigating the frequency of rubber dam use during root canal treatment, identifying influencing factors for not using it by Saudi general dental practitioners (GDPs) and endodontists. It also aimed at identifying measures that increase rubber dam usage. Methods: After obtaining an ethical approval, two pilot studies were conducted on staff members at Taibah University College of Dentistry and a group of GDPs. A final online survey was constructed comprising 17 close-ended questions divided into six categories: demographics, endodontic practice, rubber dam use, alternative isolation methods, reasons for not using rubber dam, and measures and policies that increase its usage. The survey was emailed to 375 GDPs randomly selected from the dental register and all endodontists (n = 53) working in the western province, Saudi Arabia. Data were analyzed using the Chi-square and Linear-by-Linear association tests at p ≤ 0.05. Results: The proportion of endodontists who used rubber dam (84.8 %) was significantly greater than that of GDPs (21.6 %) (p < 0.001). Significantly the highest proportion (40.5 %) did not use rubber dam because of unavailability at working place. Most rubber dam none-users (69.25 %) used a combination of other isolation means. The highest proportion of those who used rubber dam were working in the governmental sector (54.3 %). Among rubber dam users, the greatest proportion graduated from Saudi Arabia (57.8 %) compared to those graduated from Egypt (34.3 %) and Syria (22.4 %). There was a significant correlation between the patterns of rubber dam use during undergraduate training and its usage after graduation (p = 0.001). The highest proportion of participants (48.1 %) reported better undergraduate education as the most important factor that would increase rubber dam use in dental practice. Conclusions: Using of rubber dam was not common in Saudi general dental practice. Dentists must follow the recommended standards of care. Place of work and patterns of using rubber dam during undergraduate study were the most influencing factors. Better undergraduate education was the most important proposed measure to increase its usage. The combination of cotton rolls and saliva high-volume ejector or gauze was the most common alternative to rubber dam isolation. [ABSTRACT FROM AUTHOR]