Background. Acute otitis media (AOM) is often treated with antibiotics. However, initial observation is recommended. Denmark has a low use of antibiotics compared with other countries, but the total use of antibiotics has increased by 32% during the last decade, and it is important to know whether general practitioners (GPs) prescribe antibiotics according to guidelines. Objective. The aim of the study was to analyse associations between GPs' antibiotic prescribing for AOM and symptoms, diagnoses, and characteristics of children. Methods. A cohort study where GPs consecutively included 954 children between 0 and 7 years old with a new ear symptom was carried out. The GPs registered symptoms, results of otoscopy and tympanometry, together with diagnosis and treatment. Results. Fever with and without earache was statistically associated with prescribing antibiotics, and it applies to both children up to two years of age (OR: 5.89 (confidence interval (CI): 2.62–13.21) and OR: 8.13 (CI: 4.61–14.32)) and children older than two years of age (OR: 4.59 (CI: 2.55–8.25) and OR: 19.45 (CI: 6.38–59.24)). A red tympanic membrane was statistically associated with the prescribing antibiotics in both age groups (0–2 years: OR: 4.73 (CI: 2.52–8.86) and >2–7 years: OR: 3.76 (CI: 2.13–6.64)). A flat tympanometry curve was only statistically associated with prescribing antibiotics in the oldest children (OR: 2.42 (CI: 1.17–5.00)). Conclusion. This study indicates that GPs to a large degree prescribe antibiotics appropriately according to guidelines.