Beef has become a kind of important food in China because of its nutritional value perceived by consumers. With increasing consumers’ awareness and governments’ regulations on beef quality and safety, traceability is becoming a mandatory requirement in cattle/beef industry. This paper developed and evaluated a cattle/beef traceability system that integrated RFID technology with PDA and barcode printer. The system requirements, the business flow of the cattle/beef chain, and the key traceability information for the system were identified through a survey. Then a conceptual model was proposed to describe the process of traceability information acquisition, transformation and transmission along the supply chain. Finally, the system was evaluated and optimized in the sampled supply chain. The results show that the major benefits gained from the RFID-enabled traceability system are the real-time and accurate data acquisition and transmission, and the high efficiency of information tracking and tracing across the cattle/beef supply chain; the main barriers for implementing the system are the inapplicable method of inputting information, the inefficient sequence of data input and communication mechanism associated with RFID reader, and the high implementation cost.