Original research articles

Good quality Vitis RNA obtained from an adapted DNA isolation protocol

Abstract

Grapevine is a woody plant, whose high carbohydrate and phenolic compound contents usually interferes with nucleic acid isolation. After we tried several protocols for isolating RNA from the Vitis rootstock Richter- 110 (R-110) with little or no success, we adapted a method reported to be satisfactory for grapevine DNA isolation, to extract RNA. With slight protocol modifications, we succeeded to obtain polysaccharide- and phenolic-free RNA preparations from all vegetative tissues, without excessive sample handling. RNA isolated by the reported method permitted to obtain highly pure mRNA (messenger RNA) to construct a cDNA (complementary DNA) library and allowed gene transcription analysis by reverse Northern, which guarantees RNA integrity. This method may also be suitable for other plant species with high polysaccharide or phenolic contents.

Authors


Isabel Baiges

Affiliation : Unitat d'Enologia-CeRTA, Departament de Bioquimica i Biotecnologia, Facultat d'Enologia de Tarragona, Universitat Rovira i Virgili, Ramôn y Cajal, 70 43005 Tarragona, Spain


Albert Mas

Affiliation : Unitat d'Enologia-Centre de Referència en Tecnologia dels Aliments (UE-CeRTA), Departament de Bioquímica i Biotecnologia, Facultat d'Enologia de Tarragona, Universitat Rovira i Virgili, Ramón y Cajal, 70, 43005 Tarragona, Spain

amb@astor.urv.es

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