Optimization of the protocols of extraction of ADN and of the molecular marker type RAPD in Anonáceas
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Abstract
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.
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