Mangabeira (Hancornia speciosa) is a native fruit tree found mainly in the Cerrado biome and shows great economic potential due to its multiple uses; the fruits are used in agriculture, are important as a food resource, and can be consumed in natura or processed. Due to a reduction in the area of ecosystems where it occurs, mangabeira is threatened by genetic erosion in Brazil. The characterization of the genetic diversity of plants can provide the basis for strategies to protect and conserve endangered populations, like mangabeira. This study aimed to compare eight DNA extraction methods in mangabeira because the key to success is the use of a pure genomic DNA for the characterization of genetic diversity in molecular biology techniques. The quality and concentration of DNA were revealed by agarose gel electrophoresis. Polymerase chain reaction amplifications were successfully by extractions using two commercial purification kits and by the method proposed by Khanuja et al. (1999), which produced sufficient genomic DNA of good quality from leaves of H. speciosa to perform techniques involving molecular biology. The protocol described by Khanuja et al. (1999) is less expensive when compared to the commercial purification kits.