Enhanced Protocol for Isolation of Plant Genomic DNA
Muhammad Youssef, Ruby Valdez-Ojeda, José Roberto Ku-Cauich, Rosa María Escobedo-Gracia Medrano

A reproducible method for extraction of high-quality genomic DNA (gDNA) suitable for application in several PCR-based methods was developed after modifications to the Dellaporta method. Changes to the extraction buffer include the use of a higher concentration of NaCl, substitution of β-mercaptoethanol with sodium-metabisulfite, and the use of polyethylene glycol for DNA precipitation. Compared to the original method and two other protocols tested, our improved protocol resulted in the isolation of a good yield and purity of gDNA. The content of extracted DNA was spectrophotometrically evaluated, and the quality was analyzed by Amplified Fragment Length Polymorphisms (AFLP). AFLP profiles of the DNA obtained with our protocol were comparable to those of a commercial kit for plant DNA extraction. The potential of this improved method relies on its successful use with different molecular markers using gDNA extracted from fresh and frozen tissues of a variety of vascular plants, including banana in this paper, and proven in wheat, guava, sugarcane, and bean, as well as from microalgae. Therefore, the new protocol is an adequate, convenient and economical choice for use and study of various fields of genomics.

Full Text: PDF     DOI: 10.15640/jaes.v4n2a20