The primary goal of all gnotobiotic facilities is to maintain a germ-free colony until such time as protocols require associations with other organisms. As such, a well-trained and knowledgeable staff is crucial to the success of such facilities. Many facilities may differ in standard operating procedures, size, and species of animals. That being said, finding staff that correctly fit the role of a gnotobiotic technician for individual facilities is imperative. It is important to look for several key qualities in individuals during the interviewing and hiring process for any gnotobiotic position. Past laboratory animal experience, attention to detail, and organization are a few of the key concepts that aid in the gnotobiotic training process. Once an individual is hired, a detailed training process is essential to the continued success of a facility. In this presentation, we will review some basic training procedures of gnotobiotic technicians working in a mouse facility. These basic procedures range from basic animal care to facility upkeep. In addition, we will look at several additional study related procedures that are often used in gnotobiotic facilities. These will include derivation training, giving injections in isolators, special diets, and timed mating procedures for age specific studies.