Abstract

SOLANO-FLOREZ, Gina; MARQUEZ-CARDONA, María del Pilar  and  SCHULER, Ingrid. Extraction of high quality DNA of Passiflora ligularis for its analysis with molecular markers. Univ. Sci. [online]. 2009, vol.14, n.1, pp.16-22. ISSN 0122-7483.

Objective. To standardize a precise and efficient DNA isolation protocol of Passiflora ligularis. Materials and methods. Two methods of DNA extraction and two different tissues of Passiflora ligularis were evaluated in terms of purity, quality and quantity of DNA yield, as well as DNA's suitability for molecular techniques based on PCR such as RAPDs. Quantification of DNA was done using absorbance spectrophotometry at a wavelength of 260nm (A₂₆₀) with a Beckman Du ® 530 spectrophotometer. An estimate of the DNA's purity was obtained using the absorbance ratio (A₂₆₀ / A₂₈₀ nm). The variables analyzed in this study were the extraction method (A) and the tissue type (B), in order to define their influence on the purity and quantity of the DNA extracted. For the study of these variables a random design with a 2 x 2 factorial arrangement was used. Results. The average quantities of DNA obtained with the modified method of Mc Couch et al. (1988) and the modified method of Doyle & Doyle (1991) method were 778.9 µg/ml and 660.1 µg/ml respectively for dry tissue. Averages with fresh tissue were 1543.3 µg/ ml and 820.4 µg/ml respectively. Conclusion. Based upon our results we suggest the use of Mc Couch et al. (1988) method with fresh leaf tissue to obtain a high quality DNA suitable to be used with RAPDs molecular markers.

Keywords : DNA; extraction; Passiflora ligularis; RAPDs.

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