Isolation of total and polysomal RNA from plant tissues
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.
KeywordsGradient Buffer Milky Suspension Harvest Material Centrifugal Axis Fungus Schizophyllum Commune
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