ISOLATION OF GENOMIC DNAs FROM THE TROPICAL FRUIT TREES AVOCADO, COCONUT, GUAVA AND MANGO FOR PCR-BASED DNA MARKER APPLICATION
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Instituto Nacional de Ciencias Agrícolas
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With tropical fruit trees, the isolation of genomic DNA with sufficient quality for the application of PCR-based DNA marker technology very often has severe problems due to the presence of inhibitors such as polysaccharides, which inhibit enzymatic DNA processing or polyphenols as inhibitors of PCR reactions. Here, different protocols for DNA extraction and purification were tested with the four tropical fruit trees guava ( Psidium guajava L.), avocado ( Persea americana Mill.), mango ( Mangifera indica L.) and coconut ( Cocos nucifera L.). The well-established CTAB protocol of Doyle and Doyle yielded excellent DNA templates for PCR amplification with mango and coconut, but not so with guava and avocado. For these latter species, several other extraction protocols also yielded only non-satisfactory results. Modification of the CTAB method with respect to CTAB bu- ffer 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 AFLP, SSR and ISTR DNA marker analyses in guava and/or avocado.
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