Aptamers are synthetic single stranded oligonucleotides that exhibit selective binding properties to specific targets, thereby providing as powerful basis for the development of selective and sensitive (bio)chemical assays, such as biosensors. Electrochemical biosensors, utilizing aptamers as biological recognition elements, namely aptasensors, are at the forefront of current research. They exploit the combination of the unique properties of aptamers with the advantages of electrochemical detection with the view to fabricate inexpensive and portable analytical platforms for rapid detection in point-of-care (POC) applications or on-site monitoring. The immobilization of aptamers on suitable substrates is of paramount importance with the aim to retain their functionality and optimize the sensors’ sensitivity. This work describes different immobilization strategies for aptamers on the surface of semiconductor-based working electrodes, including metal oxides, conductive polymers and carbon allotropes. These are presented as platforms with tunable band gaps and different surface morphologies for the preparation of low cost, highly versatile aptasensor devices in analytical chemistry. A survey of the current literature is provided, discussing each analytical method. Future trends envisage for aptamer-based biosensing using semiconductors are also outlined.