( Simple and Rapid Isolation of Plant RN...)Simple and Rapid Isolation of Plant RNA,,, Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit

Simple and Rapid Isolation of Plant RNA Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit

Sonja Vorwerk and Barbro Patterson
Sonja Vorwerk, Max Planck Institut fr Zchtungsforschung, Carl-von-Linn Weg 10, 50829 Cologne, Germany
Phone: ++49-221-5062-441, Fax. ++49-221-5062-413, e-mail: vorwerk@mpiz-koeln.mpg.de
Barbro Patterson, Eppendorf AG, Germany Abstract

The isolation of plant RNA is a particularly challenging task. There are many protocols for the isolation of total RNA from plants that utilize solvents such as phenol or chloroform; however, their toxicity and the resulting cumbersome disposal may make alternative protocols more desirable. This application protocol demonstrates that the Eppendorf Perfect RNA Maxi Kit for eukaryotic cells is ideally suited for the isolation of total RNA from plant material, shown here with the sources Arabidopsis thaliana (dicotyledon) and Zea mays (monocotyledon).

The technology of the Perfect RNA Kit is based on cell lysis with a chaotropic guanidinium isothiocyanate solution, resulting in the rapid inactivation of cellular RNases. The RNA in the lysate is then selectively bound to a specially designed matrix and purified from DNA and other contaminants by efficient washing steps. Finally, RNA is eluted in molecular biology grade (RNase-free) water.

Materials and Methods

Arabidopsis plants, maize plants, and a maize cell culture were used for the experiments. Various amounts of starting material were used, from 110 mg (see results listed below). First, the plant material was ground very thoroughly to a fine powder in liq
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