Abstract

ALFONSO ALONSO, Yanet et al. Optimization of the protocols of extraction of ADN and of the molecular marker type RAPD in Anonáceas. cultrop [online]. 2016, vol.37, suppl.1, pp.91-98. ISSN 0258-5936.

The molecular techniques need of protocols that allow determine levels of genetic change inside the populations in different environmental conditions. So much the optimization of the isolation of the DNA, as that of the working conditions of the amplifications, they are fundamental to reach the success of the molecular analyses, therefore the present research has like objective: optimize protocols of extraction of DNA and of the molecular marker type RAPD (Random Amplified Polymorphic DNA) in Anonáceas. For the DNA extraction were used the Kit Nucleon PHYTOpure and DNeasy^® of QIAGEN. The working conditions of the protocol of amplification were fittedand changed the concentrations of DNA and of the used chokers. Followed by this, a tes was realized with 10 fatteners of the series OPH and five of the series OPA, to select more polimorphiss. The first results obtained with the Kit Nucleon PHYTOpure showed an DNA of low quality, due to the high fenolization of the vegetable material, not like that with the Kit DNeasy^® of QIAGEN, who allowed obtain an DNA of quality, purity and homogeneity to an approximate concentration of 30 ng µ L^-1. The biggest amplification products were obtained touse 3 ng µ L^-1 of choke and 2 ng µ L^-1 of DNA. Four chokers that presented major polymorphism were OPA-16, OPH-03, OPH-13 and OPH-18. The results of this research allowed optimize the working conditions of the technical RAPD for the characterization of the collection ex-situ of Anonáceas under our environmental conditions

Keywords : DNA isolation; Annonaceae; RAPD.

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