title: Optimization of DNA extraction and PCR protocols for phylogenetic analysis in Schinopsis spp. and related Anacardiaceae.
abstract: The Anacardiaceae is an important and worldwide distributed family of ecological and socio-economic relevance. Notwithstanding that, molecular studies in this family are scarce and problematic because of the particularly high concentration of secondary metabolites-i.e. tannins and oleoresins-that are present in almost all tissues of the many members of the group, which complicate the purification and amplification of the DNA. The objective of this work was to improve an available DNA isolation method for Schinopsis spp. and other related Anacardiaceae, as well as the PCR protocols for DNA amplification of the chloroplast trnL-F, rps16 and ndhF and nuclear ITS-ETS fragments. The modifications proposed allowed the extraction of 70-120 µg of non-degraded genomic DNA per gram of dry tissue that resulted useful for PCR amplification. PCR reactions produced the expected fragments that could be directly sequenced. Sequence analyses of amplicons showed similarity with the corresponding Schinopsis accessions available at GenBank. The methodology presented here can be routinely applied for molecular studies of the group aimed to clarify not only aspects on the molecular biology but also the taxonomy and phylogeny of this fascinating group of vascular plants.
authors.affiliation: Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Campo Experimental Villarino, S2125ZAA Zavalla, Santa Fe Argentina ; IICAR, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-UNR), Zavalla, Argentina.
authors.forename: Virginia Y
authors.initials: VY
authors.lastname: Mogni
authors.affiliation: SINERGIUM Biotech, Ruta Panamericana Km 38.7, 1619 Garin, Buenos Aires Argentina.
authors.forename: Mariano A
authors.initials: MA
authors.lastname: Kahan
authors.affiliation: Departamento de Ciências Biológicas, Universidade Estadual de Feira de Santana, Av. Transnordestina, s/n, Novo Horizonte, Feira de Santana, Bahia 44036-900 Brazil.
authors.forename: Luciano Paganucci
authors.initials: LP
authors.lastname: de Queiroz
authors.affiliation: Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Campo Experimental Villarino, S2125ZAA Zavalla, Santa Fe Argentina ; IICAR, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-UNR), Zavalla, Argentina.
authors.forename: José L
authors.initials: JL
authors.lastname: Vesprini
authors.affiliation: Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Campo Experimental Villarino, S2125ZAA Zavalla, Santa Fe Argentina ; IICAR, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-UNR), Zavalla, Argentina.
authors.forename: Juan Pablo A
authors.initials: JP
authors.lastname: Ortiz
authors.affiliation: Facultad de Ciencias Agrarias, Universidad Nacional de Rosario, Campo Experimental Villarino, S2125ZAA Zavalla, Santa Fe Argentina ; IICAR, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET-UNR), Zavalla, Argentina.
authors.forename: Darién E
authors.initials: DE
authors.lastname: Prado
keywords: Anacardiaceae; DNA isolation; Phylogeny; Schinopsis; Sequencing