Standardization of a protocol for obtaining genomic DNA for the quantification of 5mC in epicormic outbreaks of Tectone grandis L.
Journal Title: Biotecnología Vegetal - Year 2008, Vol 8, Issue 1
Abstract
The present investigation was carried out with the objective of defining an extraction and purification method that it provided deoxyribonucleic acid (DNA) appropriate to determine the percentage of 5mC in the genomic DNA of epicormics buds of Tectona grandis L. During the standardization of the protocol four methods were compared: 1 - classic based on saline shock solution with CTAB (hexadecil trimetil ammonium bromide), 2 - Kit of extraction of DNA plants DNeasy Plant Mini Kit (QIAGEN) according to the protocol recommended by the maker, 3 - Kit of extraction of DNA plants DNeasy Plant Mini Kit (QIAGEN) modified with phenol employment without silicon columns, 4 - Kit of extraction of DNA plants DNeasy Plant Mini Kit (QIAGEN) modified with phenol and additionally silicon columns employment. The samples extracted with CTAB method, generated totally non valid electroferograms for the nucleosides presence. Valid electroferograms were obtained only when the DNA was extracted and purified with the Kit DNeasy Plant Mini Kit (QIAGEN) modified with phenol and additionally using silicon columns, with this protocols only desoxinucleosides is obtained being verified the high purity of the samples. We described an extraction and purification method that provided genomic DNA of teak based on the phenol employment and purification with columns of SiO 2 , appropriate to determine in a precise way the percentage of 5mC in the genomic DNA of epicormics buds of Tectona grandis L.
Authors and Affiliations
Elisa Quiala , Luis Valledor , Rodrigo Hazbun , Raúl Barbón , Manuel de Feria , Maité Chávez , Roberto Rodríguez
Keywords
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