A New Method for Rapid Extraction of High Quality RNA from Recalcitrant Tissues of Grapevine

来自 EBSCO

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摘要：

A quick, inexpensive, and reliable protocol for the extraction of RNA from grapevine berry skins containing large quantities of polyphenols, procyanidins, and polysaccharides is described. The method involves an extraction step in the presence of ribonuclease inhibitors and compounds that compete with vacuolar contaminants for binding to RNA. After extraction with organic solvents, RNA is bound to a fibrous cellulose matrix and processed to eliminate the remaining contaminants and ribonucleases. Following this method, highly stable RNA, sufficiently pure for northern hybridizations and enzymatic processing, may be obtained from as little as 200 mg of starting amounts of fresh material and without multiple, time consuming precipitations or ultracentrifugation steps. This procedure may also prove useful for extracting RNA from recalcitrant tissues of other plant species. Abbreviations: ATA, aurintricarboxylic acid; CF11, cellulose fibrous medium (type 11); PVPP, polyvinylpolypyrrolidone; RT room temperature; VRC, vanadyl ribonucleoside complex.

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关键词：

RAPID METHODS, polyphenols, genetica, polisacaridos, methods, flavonoide, polyholoside, inhibiteur d'enzyme, genetique, italie, methode, tecnicas de aislamiento, genetics, italia, fruits, purification, frutas, BINDING SITES, inhibidores de enzimas, tecnicas analiticas, arn, italy, ribonuclease, polysaccharides, metodos, rna, vitis vinifera, polifenoles, ribonucleasas, technique analytique, polyphenol, analytical methods, ribonucleases, technique de l'isolement, CYANIDIN, METHODOLOGY, flavonoides, GENETIC ANALYSIS, isolation techniques, flavonoids, enzyme inhibitors, purificacion

DOI：

10.1023/A:1007433910213

被引量：

147

年份：

1998