A new method for rapid extraction of high quality RNA from recalcitrant tissues of grapevine

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 wi...

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Bibliographic Details
Published in:Plant molecular biology reporter 1998-03, Vol.16 (1), p.61
Main Authors: Geuna, F. (Universita di Milano, Milano, Italy.), Hartings, H, Scienza, A
Format: Article
Language:English
Subjects:
Abbreviations
ANALYTICAL METHODS
ARN
Aurintricarboxylic acid
BINDING SITES
Cellulose
Cellulose fibers
Contaminants
CYANIDIN
ENZYME INHIBITORS
FLAVONOIDE
FLAVONOIDES
FLAVONOIDS
FRUITS
FRUTAS
GENETIC ANALYSIS
GENETICA
GENETICS
GENETIQUE
Hybridization
INHIBIDORES DE ENZIMAS
INHIBITEUR D'ENZYME
ISOLATION TECHNIQUES
ITALIA
ITALIE
ITALY
METHODE
METHODOLOGY
METHODS
METODOS
Organic solvents
Plant extracts
Plant species
Plant tissues
POLIFENOLES
POLISACARIDOS
POLYHOLOSIDE
POLYPHENOL
POLYPHENOLS
POLYSACCHARIDES
Polyvinylpolypyrrolidone
Procyanidins
PURIFICACION
PURIFICATION
RAPID METHODS
RIBONUCLEASAS
RIBONUCLEASE
Ribonuclease inhibitors
RIBONUCLEASES
Ribonucleic acid
RNA
Saccharides
Skin
TECHNIQUE ANALYTIQUE
TECHNIQUE DE L'ISOLEMENT
TECNICAS ANALITICAS
TECNICAS DE AISLAMIENTO
Ultracentrifugation
VITIS VINIFERA
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Description
Summary: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.
ISSN:0735-9640
1572-9818
DOI:10.1023/A:1007433910213