Midazolam is increasingly used in reptiles as part of a multidrug induction for general anesthesia. Studies of its use as a sole sedative are limited to a few reports in turtles and crocodiles. A retrospective review was carried out to assess the clinical efficacy of midazolam as a sedative in reptiles at the National Aquarium, Baltimore, Maryland. Medical procedures from 48 events involving 34 individuals were included in the review. Information obtained included species, age, sex, body weight, type of procedure performed, dosage of midazolam administered, use of a concurrent analgesic, time to effect, use of a reversal agent, and efficacy of sedation (acceptable vs. unacceptable). Midazolam was used in a wide variety of small- to medium-sized turtles, lizards, and snakes. Diagnostic imaging (e.g., radiography) and minor surgical procedures—in combination with local anesthetics (e.g., distal digit amputation)—were the most common procedures performed. The median dosage of midazolam was 0.3 mg/kg (range 0.1–1.0 mg/kg). Acceptable sedation was achieved in 80% of events. Midazolam can provide acceptable sedation to facilitate minor procedures in a variety of reptiles using dosages that are lower than previously reported in the literature.