Isolation of genomic DNAs from the tropical fruit trees avocado, coconut, guava and mango for PCR-based DNA marker application.
Ramírez I. M., Rodríguez N. N., Valdés-Infante J., Capote M., Becker D., Rohde W.
Author Affiliation: Centro de Aplicaciones Tecnológicas y Desarrollo Nuclear (CEADEN), Miramar, Playa, Ciudad de La Habana, Cuba.
Cultivos Tropicales 25 : 33-38
Abstract : With tropical fruit trees, the isolation of a sufficient quality of genomic DNA for use in PCR-based DNA marker technology very often poses severe problems due to the presence of inhibitors such as polysaccharides, which inhibit the enzymatic DNA processing or polyphenols as inhibitors of PCR reactions. Here, different protocols for DNA extraction and purification were tested using 4 tropical fruit trees viz., guava (Psidium guajava), avocado (Persea americana), mango (Mangifera indica) and coconut (Cocos nucifera). The well-established CTAB protocol of Doyle and Doyle yielded excellent DNA templates for PCR amplification for mango and coconut, but not so with guava and avocado. For these latter species, several other extraction protocols also yielded non-satisfactory results. Modification of the CTAB method with respect to CTAB buffer composition and in combination with reversible adsorption to NucleoSpin columns alleviated the problems encountered with the genomic DNA of both species. The quality of DNA prepared by this procedure allowed amplified fragment length polymorphism, simple sequence repeat and inverse sequence-tagged repeat DNA marker analyses in guava and/or avocado.