RNA isolation from loquat and other recalcitrant woody plants with high quality and yield

Please use this identifier to cite or link to this item: http://hdl.handle.net/10045/35691
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Title: RNA isolation from loquat and other recalcitrant woody plants with high quality and yield
Authors: Morante Carriel, Jaime | Sellés Marchart, Susana | Martínez Márquez, Ascensión | Martínez Esteso, María José | Luque Romero, Ignacio | Bru-Martinez, Roque
Research Group/s: Proteómica y Genómica Funcional de Plantas
Center, Department or Service: Universidad de Alicante. Departamento de Agroquímica y Bioquímica
Keywords: RNA isolation | Fruits | Polyphenols | Polysaccharides | Woody plants
Knowledge Area: Bioquímica y Biología Molecular
Issue Date: 17-Feb-2014
Publisher: Elsevier
Citation: Analytical Biochemistry. 2014, Accepted Manuscript, Available online 17 February 2014. doi:10.1016/j.ab.2014.02.010
Abstract: RNA isolation is difficult in plants that contain large amounts of polysaccharides and polyphenol compounds. To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A260/280 ratios ranged from 1.90 to 2.04; A260/230 ratios were >2.0), but also of high yield (up to 720 μg on average [CV 21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, bruising), leaf, root, flower, stem and bud; quince fruit and root; grapevine cells in liquid culture; and, rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression and function.
Sponsor: This work has been supported by Spanish Ministry of Foreign Affairs and Cooperation (A/8823/07) and (B/017931/08), the Spanish Ministry of Science and Innovation (BIO2011-29856-C02-02) and European Funds for Regional Development (FEDER).
URI: http://hdl.handle.net/10045/35691
ISSN: 0003-2697 (Print) | 1096-0309 (Online)
DOI: 10.1016/j.ab.2014.02.010
Language: eng
Type: info:eu-repo/semantics/article
Peer Review: si
Publisher version: http://dx.doi.org/10.1016/j.ab.2014.02.010
Appears in Collections:INV - Proteómica y Genómica Funcional de Plantas - Artículos de Revistas

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