Comparison of three genomic DNA extraction methods from soursop leaves (Annona muricata L.)

Abstract

The new molecular biology techniques require high-quality DNA. Most of the DNA extraction methods rely on commercially available kits and/or the cetyltrimethylammonium bromide (CTAB) method. Up to date, there is a lack of molecular studies in soursop (Annona muricata L.) and no DNA extraction protocol has been reported. Therefore, the development of a method to extract high-quality soursop DNA is necessary. In this study, we compared three methods to isolate genomic DNA from lyophilized soursop leaves. The DNA was extracted using the MO BIO Laboratories Inc. PowerPlantR Pro DNA Isolation Kit and two CTAB-methods with some modifications. The parameters evaluated were concentration, purity, and integrity. Besides, the maturase K (matK) gene was amplified by polymerase chain reaction (PCR) to test the effectiveness of DNA. Our results showed higher DNA concentration and purity using the Sagahi-Maroof method in comparison with the Doyle & Doyle and the molecular kit. Therefore, the combination of 3% polyvinylpyrrolidone (PVP) and 3% CTAB in the lysis buffer improved the quality and concentration of the DNA extracted from soursop leaves. Further, the matK gene with a size of 796 bp was successfully amplified by PCR from the DNA isolated with the Sagahi-Maroof method in all samples tested. In conclusion, the Sagahi-Maroof CTAB-method with modifications was the most efficient method to extract high-quality DNA, which will serve for future molecular studies.