Isolation of total and polysomal RNA from plant tissues

  • Sacco De Vries
  • Harry Hoge
  • Ton Bisseling

Abstract

Most plant material contains relatively high levels of RNase activity which is normally located in the vacuoles. During the RNA extraction procedure RNA should be protected against this endogenous RNase. In this chapter we describe two procedures for the isolation of RNA. In both procedures a high pH of the extraction buffer and the presence of a chelating agent (EDTA and EGTA respectively) are used to prevent RNA degradation. In addition, during the isolation of total RNA a detergent (SDS) is used and the pulverized material is directly thawed in a mixture of phenol and extraction buffer (denaturing the RNase). For both RNA extraction procedures we found that the addition of RNase inhibitors was unnecessary, thereby omitting complicated and expensive buffers.

Keywords

Gradient Buffer Milky Suspension Harvest Material Centrifugal Axis Fungus Schizophyllum Commune 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Kluwer Academic Publishers, Dordrecht 1989

Authors and Affiliations

  • Sacco De Vries
    • 1
  • Harry Hoge
    • 2
  • Ton Bisseling
    • 1
  1. 1.Department of Molecular BiologyAgricultural UniversityWageningenNetherlands
  2. 2.Department of BiochemistryState University LeidenLeidenNetherlands

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