Medicinal plants have been used for the treatment of diseases and its demand is expanding in both developed and developing countries. The extensive consumption to meet demand:supply ratio exerts a heavy strain on the existing resources. This subsequently led to adulteration and substitution. The DNAbased method is the most widely adopted for the determination of adulteration in herbal products.Quality DNA extraction is regarded as the prerequisite for the application of DNA-based techniques in herbal plant authentication. Various protocols for DNA isolation could only be applied to a small number of species and were often expensive and time-consuming. Furthermore, herbal products undergo various conditions and are mixed with different substances, these factors affect the integrity of the DNA making it di cult to isolate. In this study, a simple, cost-effective and rapid CTAB-optimized DNA extraction protocol was developed which successfully isolated quality DNA from different samples of medicinal plants and their respective adulterants without using liquid nitrogen. Quality DNA was also isolated from different herbal products of reputed companies. The protocol was also validated in different herbarium samples and quality DNA has been obtained. DNA quality of both plants and herbal drugs was measured by A260/A280 ranging between 1.3-1.8. Thus, the developed protocol can be adapted to medicinal plants and herbal drugs quality DNA isolation and authentication as well as herbarium samples for molecular identi cations. The isolated DNA was found to have suitable quality and su cient quantity for ISSR-PCR, SCoT-PCR and restriction digestion reaction, advocating the protocol is best suitable for different genomics downstream applications.