Optimized method for dna extraction and PCR amplification in aroeira tree

Jailson do Nascimento Silva; Amanda Camila dos Santos Linhares; Dario Abel Palmieri; Marcones Ferreira Costa; Gisele Holanda de Sá; Maria Fernanda da Costa Gomes; Michelli Ferreira dos Santos; Lidiane de Lima Feitoza; Sérgio Emílio dos Santos Valente

doi:10.14393/BJ-v39n0a2023-62577

Authors

Jailson do Nascimento Silva Instituto Federal do Piauí
Amanda Camila dos Santos Linhares Universidade Federal do Piauí
Dario Abel Palmieri Universidade Estadual Paulista https://orcid.org/0000-0003-0841-6257
Marcones Ferreira Costa Universidade Federal do Piauí https://orcid.org/0000-0001-8210-2673
Gisele Holanda de Sá Universidade Federal do Piauí https://orcid.org/0000-0002-0687-9314
Maria Fernanda da Costa Gomes Universidade Estadual do Piauí https://orcid.org/0000-0002-1089-8593
Michelli Ferreira dos Santos Universidade Federal do Piauí https://orcid.org/0000-0001-7668-0864
Lidiane de Lima Feitoza Universidade Federal do Piauí
Sérgio Emílio dos Santos Valente Universidade Federal do Piauí https://orcid.org/0000-0003-2953-7330

DOI:

https://doi.org/10.14393/BJ-v39n0a2023-62577

Keywords:

CTAB, DNA isolation, Myracrodruon urundeuva.

Abstract

Molecular markers are important tools in the characterization of plant genetic diversity and can provide support for conservation strategies for endangered populations. The different molecular techniques involve the evaluation of many individuals; therefore, it is crucial to have fast, efficient, and inexpensive methods for DNA extraction. Given the importance of the Aroeira (Myracrodruon urundeuva Fr. All.) it is pertinent to optimize a protocol that allows the obtainment of intact and pure DNA, aiming to assist conservation strategies for this species that is threatened with extinction. Thus, this study aimed to compare five DNA extraction methods: Dellaporta et al. (1983), Doyle and Doyle (1987) modified, Ferreira and Grattapaglia (1995), Romano and Brasileiro (2015), and Khanuja et al. (1999) and optimize the most efficient protocol for M. urundeuva. The modified DNA extraction protocol proposed by Doyle and Doyle (1987), using 100 mg of leaf tissue and 6 µl of β-mercaptoethanol was the protocol that presented the sharpest bands after DNA electrophoresis and after the reactions of amplification employing Polymerase Chain Reaction (PCR). Therefore, it is suggested to use the protocol described by Doyle and Doyle (1987) modified for the extraction of DNA from young M. urundeuva leaves to carry out techniques involving molecular markers.

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