Abstract: This study was conducted to develop a rapid and efficient protocol for extracting high quality DNA from Pistachio trees suitable for PCR and molecular studies. Genomic DNA was extracted from 12 Pistachio trees using modified QIAGEN DNeasy Plant Mini Kit. The results showed that the modified protocol successfully produced a sufficient amount of DNA with high quality, which was highly confirmed by the purity index values of DNA samples (1.45 to 2.01). In conclusion, the modified protocol can produce high quality DNA from Pistachio trees suitable for PCR studies such as RAPD and AFLP and it can be easily adjusted for other Pistacia species.