Molecular typing of pathogenic bacteria plays a crucial role in the food industry, enabling the detection, tracing, and control of foodborne pathogens such as Listeria monocytogenes. Among them, Pulsed Field Gel Electrophoresis (PFGE) plays an important role in the epidemiological characterization of these pathogens, identifying clonal relationships between isolates from food, environment, and clinical sources. This method has proven useful in outbreak investigation and understanding the pathogen persistence in food processing environments, contributing to the design of specific control strategies. This chapter details a step-by-step PFGE protocol for Listeria monocytogenes based on the standardized PulseNet protocol, providing detailed explanations of critical parameters to ensure accuracy and reproducibility.