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An Improved Method to Isolate Mitochondrial RNA from Green Plant Tissue

  • Fei Ye
  • Ralf Reski
Protocol
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Part of the Springer Protocols Handbooks book series (SPH)

Abstract

In plant cells, mitochondrial RNA (mtRNA) constitutes about only 1% of the total RNA. From this, most RNAs are ribosomal RNAs. Thus, isolation of high purified mtRNA is necessary, not only for construction of a mitochondrial cDNA library, but also for the analysis of plant mitochondrial transcription. Several methods have been frequently used for isolation of plant mtRNA (1, 2, 3). However, these mtRNA preparations may be heavily contaminated by chloroplast RNA (cpRNA), especially when mtRNA is isolated from green leaves (1,4). It is believed that the cpRNA sticks to the mitochondrial membrane and therefore persists after gradient purification of mitochondria. Although micrococcal nuclease would be the enzyme to remove the non-mtRNA from mitochondrial membranes prior to lysis of mitochondria, treatments with micrococcal nuclease for the mtRNA isolation from green leaves have not been effective (4).

Keywords

Potassium Acetate Micrococcal Nuclease Guanidinium Thiocyanate Sucrose Gradient Centrifugation Guanidine Thiocyanate 
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

© Humana Press Inc., Totowa, NJ 2000

Authors and Affiliations

  • Fei Ye
    • 1
  • Ralf Reski
    • 2
    • 3
  1. 1.Department of BiologyMassachusetts Institute of TechnologyCambridge
  2. 2.Freiburg University Institute Biology IIFrieburg/BreisguaGermany
  3. 3.OR Institute for General BotanyHamburgGermany

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