Deoxyribonucleic acid (DNA) plays an extremely important role in storing genetic messages that are used for cellular functions in living organisms. However, the process of replicating these messages is not free from errors as internal and external factors can contribute to the damages of DNA strands. These damages known as lesions are problematic since they can lead to mutagenesis and carcinogenesis after accumulation for a certain period of time. Thankfully, cells are able to synthesize various DNA glycosylases to target different lesions, repairing and restoring damaged DNA to its original state. This review article aims to explore the variety of assays used to evaluate the efficiency of these glycosylases, mainly focusing on uracil DNA glycosylase, thymine DNA glycosylase, and 8-oxoguanine DNA glycosylase since they are deployed to the sites of damages much more often than other repair enzymes. The progress in the development of DNA glycosylase assays is reviewed with an emphasis on the configuration and operating mechanism of the assays and with some discussion on challenges and prospects. It is hoped that this article will provide an introductory overview and the current standing of DNA glycosylase assays, opening up new perspectives towards the development DNA glycosylase assays for uses in molecular biology and at point-of-care. (C) 2016 Elsevier B.V. All rights reserved.