This chapter focuses on the comprehensive overview of design, simulation, manufacturing, and scale-up techniques used in microfluidic chip fabrication. The chapter begins with an introduction to the lab-on-a-chip approach and explains the chip design and simulation methods. It also highlights the various software tools and methodologies used to optimize the chip performance, including computational fluid dynamics simulations. The next section focuses on prototyping techniques for translating designs into physical devices. Mainly, four crucial methods are addressed in detail: polydimethylsiloxane soft lithography, laser machining of polymers, hot embossing and 3D printing (especially Digital Light Processing). The benefits and drawbacks of each method for specific applications in microfluidic chip fabrication are detailed in this chapter. Different procedures related to the scale-up process are explained. These include electroforming, micromachining, tooling correction, micro injection molding, bonding techniques, surface treatment methods, and reagent storage strategies. Additionally, the integration of sensors and electrodes into the microfluidic chip is explored, presenting the importance of seamless integration for accomplishing enhanced performance. Quality control and performance validation are addressed at the end of the chapter. This book chapter serves as a valuable resource for researchers, engineers, and scientists working in the field of microfluidic chip fabrication.