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Full-Length Gene Enrichment by Using an Optimized RNA Isolation Protocol in Bixa orellana Recalcitrant Tissues

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Abstract

A reliable protocol is described for isolation of large full-length cDNA from Bixa orellana mature tissues containing large quantities of pigments, phenols, and polysaccharides. This protocol involves the optimization of a commercial RNA extraction protocol in combination with a long distance reverse transcript PCR protocol. The principal advantages of this protocol are its high RNA yield and quality. The resulting RNA is suitable for RNA expression evaluation and production of large, full-length cDNA. This is the first time RNA has been isolated from all mature tissues in the tropical perennial plant B. orellana and has been proved viable for downstream applications, especially important for molecular biology studies on this economically important pigment-producing plant.

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Acknowledgments

This research was supported by the Consejo Nacional de Ciencia y Tecnología (CONACyT) through projects 46541; IFS F/2932-2 and UC-Mexux. NLRA was supported by a post graduate scholarship from the CONACyT (Grant No. 196432).

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Authors and Affiliations

  1. Centro de Investigación Científica de Yucatán, Calle 43 No. 130, Col. Chuburná de Hidalgo, 97200, Merida, Yucatan, Mexico

    N. L. Rodríguez-Ávila, M. L. Aguilar-Espinosa & R. Rivera-Madrid

  2. Centro de Biotecnología Genómica del Instituto Politécnico Nacional, Blvd. del Maestro s/n, Col. Narciso Mendoza, 88710, Reynosa, Tamaulipas, Mexico

    J. A. Narváez-Zapata

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  1. N. L. Rodríguez-Ávila
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  2. J. A. Narváez-Zapata
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  3. M. L. Aguilar-Espinosa
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  4. R. Rivera-Madrid
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Correspondence to R. Rivera-Madrid.

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Rodríguez-Ávila, N.L., Narváez-Zapata, J.A., Aguilar-Espinosa, M.L. et al. Full-Length Gene Enrichment by Using an Optimized RNA Isolation Protocol in Bixa orellana Recalcitrant Tissues. Mol Biotechnol 42, 84–90 (2009). https://doi.org/10.1007/s12033-008-9138-4

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