Simple and Rapid Isolation of Plant RNA,,, Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit
Simple and Rapid Isolation of Plant RNA Using the Eppend...The isolation of plant RNA is a particularly challenging task. There ...The technology of the Perfect RNA Kit is based on cell lysis with a ch... Materials and Methods ...Arabidopsis plants maize plants and a maize cell culture were used ...
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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