This paper describes the implementation of Design of Experiment (DOE) for RF and Microwave course. The objective is to demonstrate the use DOE in investigative laboratory activity for 3rd year students in RF and Microwave Course. Detailed explanation on the proposed framework, student activity, formulation, and learning process were outlined in an open-ended cooperative laboratory activity. The scenario requires students to determine which Microstrip Transmission Line parameter significantly affects the performance of the transmission line due to a AE 5% physical variation in the parameter from its nominal value. The outcome from the study outlined the basic procedure to apply DOE as a learning process for determining factors influencing the output functional performance. ß 2015 Wiley Periodicals, Inc. Comput Appl Eng Educ 23:658-666, 2015; View this article online at wileyonlinelibrary.com/journal/cae;