In Brazil, it is estimated that at least 10% of the native flora, which corresponds to about 4 or 5 thousand plant species, are used for food purposes. Among these species,Pereskia aculeataMill, known as ora-pro-nóbis and belonging to the Cactaceae family, stands out. It is considered a Unconventional Food Plant (UFP) and a vegetable of traditional use, playing an important role in the diet and economy of rural and urban families. Due to their high nutrient content, they have both economic and social significance, being a target of interest for scientific research. Despite the potential for human use of this species, there is a lack of knowledge about its diversity and genomic structure, resulting in few genetic and molecular studies that can contribute to the conservation and improvement of this species in general. It is essential to acquire information about its genomic structure, as well as to carry out studies related to the comparison, selection and optimization of protocols for obtaining quality DNA and in sufficient quantity, in order to guarantee the success of molecular and evolutionary analyses. The aim of this research was to evaluate qualitatively and quantitatively methods of isolation of genomic DNA fromPereskia aculeataMill. Significant variations were observed in the DNA extracted and in the purity observed between the different protocols tested, with the isolation by the protocol described by Sambrook et al. (1989) being the most efficient in terms of concentration and purity, followed by the modified method proposed in this work. This work is a pioneer in comparing methods for isolating DNA fromPereskia aculeataMill. in terms of quality and quantity. In addition, it was found that there is still a scarcity of molecular studies on this species, both worldwide and in Brazil, which indicates the existence of a promising gap for the development and realization of scientific research in this field.