This study aims to address the challenge of low-cost hardware implementation of a combined backstepping with fractional order PID (FOPID) controller for mobile robots in real-time applications. Moreover, this work proposes a self-designed mobile robot prototype that is easy to realize, low in cost, spares time, and reduces human effort. This robot platform was equipped with two DC motors with quadratic encoders and two passive wheels, controlled by an Arduino mega, where the software code was developed in the Matlab-Simulink environment, using Simulink support package for Arduino. Four case studies were conducted to demonstrate the effectiveness of the suggested methodology. Experimental results demonstrate improved trajectory tracking performance with less tracking error and smooth control efforts, and is capable of handling trajectories with continuous and non-continuous gradients.