This chapter provides a comprehensive guide to plant tissue culture, focusing on the essential equipment, types of media, and detailed practical protocols necessary for successful cultivation. It begins with an overview of the critical equipment required for plant tissue culture. The chapter then explores into the various types of media used in plant tissue culture, such as Murashige and Skoog (MS) and Gamborg B5, detailing their composition and preparation methods. Following the foundational knowledge of equipment and media, the chapter offers step-by-step practical protocols for media preparation and the surface sterilization of explants. These protocols are designed to ensure aseptic conditions and optimal growth environments for plant tissues. Each procedure is explained with precision, providing clear instructions on the materials required, preparation steps, and critical considerations to avoid contamination and ensure successful culture initiation. By integrating theoretical concepts with hands-on practical instructions, this chapter serves as an invaluable resource for students, researchers, and practitioners in the field of plant tissue culture. It aims to bridge the gap between academic knowledge and practical application, enabling readers to confidently set up and conduct their own plant tissue culture experiments.