We evaluated antibiotic prescription practices during root canal treatments among general dentists in private dental clinics in Al-Madinah Al Munawarah, Saudi Arabia. Methods: A self-administered, questionnaire about antibiotic used during root canal treatment was distributed to 75 randomly selected general dental practitioners working in private dental clinics in Al-Madinah Al-Munawarah, Saudi Arabia, between March and April 2016. The questionnaires were collected one week later. To compare results of the collected data, Chi-square test was used. Results: The results revealed that 60% of the dentists prescribed amoxicillin with clavulanic acid as the first choice treatment for endodontic pathosis. Clindamycin (51.6%) was the first choice for patients who were allergic to penicillin. Forty-five percent of the general practitioners prescribed antibiotics for 5 days. Approximately 83.3% of general practitioners prescribed antibiotics for acute apical abscesses. Prophylactic antibiotics were prescribed for cases with a history of infective endocarditis (65.5%), non-controlled diabetes (60.3%), placement of a prosthetic joint in the previous 2 years (46.6%), congenital heart disease (36.2%), and kidney dialysis shunts (34.5%). Conclusion: This study reveals antibiotic abuse in endodontic treatment practice in private dental clinics in Al-Madinah Al Munawarah, Saudi Arabia. General dental practitioners are lacking knowledge regarding the prescription of antibiotics in endodontic treatment and situations requiring prophylactic antibiotics.