DNA EXTRACTION METHODOLOGIES IN THE VINE AND RAPD ANALYSIS FOR AFLP1

Abstract

Genus Vitis is of great economic importance worldwide and is taking this subject of several genetic studies. The objective of this study is to assess the quality of DNA extracted from young leaves of 19 grape cultivars, using six extraction methods - Saghay-Maroof, Soleman and Allard (1984); Shillito and Saul (1988); Doyle and Doyle (1991); Lodhi et al (1994).; This, Cuisset and Boursiquot (1997) and Kuo Line (1998) - for RAPD and AFLP analysis. The DNA obtained was quantified by spectrophotometry and quality was assessed by electrophoresis on 0.8% agarose gel stained with ethidium bromide (0.05 g / ml). For RAPD-PCR analysis we used seven primers (B12, B13, C12, D6, E10, J5 and UBC 301). After electrophoresis on a 1.5% agarose gel, the results were visualized Gel-doc. By the extraction described by Shilito and Saul (1988) no DNA was obtained in any of the cultivars, the same happened when using the Methodology of Line Kuo (1998) when the cultivars were rootstocks. Despite other methodologies make possible the extraction of sufficient DNA, these DNAs showed up degraded or not amplified, with the exception of Lodhi et al. (1994), for RAPD-PCR analysis, and Doyle and Doyle (1991) for AFLP analysis.