Evaluation of DNA extraction methodologies from recalcitrant plants
DOI:
https://doi.org/10.5377/torreon.v9i24.9723Keywords:
DNA, extraction, commercial kits, protocols, recalcitrantAbstract
Obtention of high-quality purified deoxyribonucleic acid (DNA) is key for the subsequent application of molecular techniques during the characterization of species under study. In this regard, plant extracts can be challenging due to its complex metabolites composition and to the fact that often these chemicals concentration differs across varieties and even samples. This is, for instance, the case for recalcitrant samples of cocoa varieties (Theobroma cacao) and coffee (Coffea arabica), where DNA loss can occur during sample maceration due to phenolization. In this research, three commercial kits, already known by producing excellent recoveries for non-recalcitrant varieties, were evaluated. Additionally, the modification of traditional protocols [Doyle & Doyle (1987) and Doyle & Doyle (1990)] was also evaluated. Results obtained confirm the effect of variety and species on DNA extraction and for some species modified methods may yield DNA recovery and purity higher than those obtained by commercial kits.
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