Despite the importance in conservation and breeding purposes, molecular studies using Cerrado plant species are still rare, mainly because of their high amounts of secondary compounds, impeding DNA extraction to downstream applications, such as PCR amplification. To date, the DNA extraction methods described are sometimes inadequate for these species, expensive, time-intensive and/or use very toxic reagents. Here, we present a simple and effective method, based on SDS and Triton X-100, to obtain high DNA quality and quantity from Cerrado plant species for molecular biological techniques. The DNA obtained by our protocol was free of contaminants and excellent for enzymatic restriction and PCR amplification. The concentration of extracted DNA across all tested species ranged from 156 to 1166 ng µL-1 (1.56-11.66 µg g-1 of dry tissue), with an A260/A280 ratio from 1.78 to 1.92. The new DNA extraction protocol described here provides high DNA quality and quantity from Cerrado plant species in a fast, simple and less toxic way. Thus, the use of our method will allow ecologists, geneticists and breeders to rapidly obtain high-quality and -quantity DNA from Cerrado plant species for any molecular biology study.