Skip to main content

Extraction of RNA from Plants

  • Protocol

Part of the book series: Methods in Molecular Biology ((MIMB,volume 4))

Abstract

Current research into the structure and function of plant genes involves the application of many elaborate techniques for gene cloning and analysis. The isolation of pure, intact plant mRNA is required at many stages in this process, e.g., for generation and screening of cDNA clones, for characterization and mapping of cloned genes, and for the study of gene expression. The isolation of RNA from plants, however, often presents more problems than many of the sophisticated procedures used subsequently. Not only is there the problem of RNA degradation by endogenous and exogenous nucleases (common to RNA isolation from any organism), there is also the particular problem of extracting RNA from plant material that may be rich in starch, pectins, phenolics, and various other secondary products. Many of these plant products are not removed during conventional phenol extraction procedures, such as the one described previously in this series (1). Clearly the problems encountered in obtaining clean, undegraded RNA from any particular plant tissue will vary according to the level of nuclease activity and the range of possible contaminants, and it might be expected that different solutions would be required in each case.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   169.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   219.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Slater, R.J. (1985) The extraction of total RNA by the detergent and phenol method, in Methods in Molecular Biology Vol. 2 (Walker, J., ed.) Humana, Clifton, New Jersey.

    Google Scholar 

  2. Grierson, D., Slater, A., Spiers, J., and Tucker, G. A. (1985) The appearance of polygalacturonase mRNA in tomatoes: One of a series of changes in gene expression during development and ripening. Planta 163, 263–271.

    Article  CAS  Google Scholar 

  3. Speirs, J., Brady, C.J., Grierson, D., and Lee, E. (1984) Changes in ribosome organisation and messenger RNA abundance in ripening tomato fruits. Aust. J. Plant Physiol 11, 225–233.

    Article  CAS  Google Scholar 

  4. Palmiter, R.D. (1974) Magnesium precipitation of ribonucleoprotein complexes. Expedient techniques for the isolation of undegraded polysomes and messenger ribonucleic acid. Biochemistry 13, 3606–3615.

    Article  PubMed  CAS  Google Scholar 

  5. Mozer, T.J. (1980) Partial purification and characterisation of the mRNA for a-amylase from barley aleurone layers. Plant Physiol. 65, 834–837.

    Article  PubMed  CAS  Google Scholar 

  6. Slater, A., Maunders, M.J., Edwards, K., Schuch, W., and Grierson, D. (1985) Isolation and characterisation of cDNA clones for tomato polygalactouronase and other ripening-related proteins. Plant Mol. Biol. 5, 137–147.

    Article  CAS  Google Scholar 

  7. Maunders, M.J., Slater, A., and Grierson, D. (1985) Generation and Use of cDNA Clones for Studying Gene Expression, in Molecular Biology and Biotechnology (Walker, J.M. and Gingold, E.M., eds.) Royal Society of Chemistry, London.

    Google Scholar 

  8. Slater, A. (1987) Hybrid-Release Translation, in Methods in Molecular Biology, Vol. 4 (Walker, J., ed.) Humana, Clifton, New Jersey.

    Google Scholar 

  9. Kitos, P.A., Saxon, G., and Ames, H. (1972) The isolation of polyadenylate with unreacted cellulose. Biochem. Biophys. Res. Comm. 47, 1426–1437.

    Article  PubMed  CAS  Google Scholar 

  10. Holley, R.W., Apgar, J., and Merrill, S.H. (1961) Evidence for the liberation of a nuclease from human fingers. J. Biol. Chem. 236, PC42.

    PubMed  CAS  Google Scholar 

  11. Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982) Molecular Cloning, A Laboratory Manual Cold Spring Harbor Laboratories, Cold Spring Harbor, New York.

    Google Scholar 

  12. Slater, R.J. (1985) Purification of Poly (A)-Containing RNA by Affinity Chromatography, Methods in Molecular Biology Vol. 2 (Walker, J., ed.) Humana, Clifton, New Jersey.

    Google Scholar 

  13. Davies, E., Larkins, B.A., and Knight, R.H. (1972) Polyribosomes from peas. An improved method for their isolation in the absence of ribonuclease inhibitors. Plant Physiol. 50, 581–584.

    Article  PubMed  CAS  Google Scholar 

  14. Gray, J.C. (1974) The inhibition of ribonuclease activity and the isolation of polysomes from leaves of the French bean, Phaseolus vulgaris. Arch. Biochem. Biophys. 163, 343–348.

    Article  CAS  Google Scholar 

  15. Blackburn, P., Wilson, G., and Moore, S. (1977) Ribonuclease inhibitor from human placenta. J. Biol. Chem. 252, 5904–5910.

    PubMed  CAS  Google Scholar 

  16. Smith, C.J.S., Slater, A., and Grierson, D. (1986) Rapid appearance of an mRNA correlated with ethylene synthesis encoding a protein of molecular weight 35,000. Planta 168, 94–100.

    Article  CAS  Google Scholar 

  17. Loomis, W.D. (1974) Overcoming problems of phenolics and quinones in the isolation of plant enzymes and organelles. Meth. Enzymol. 31, 528–544.

    Article  PubMed  CAS  Google Scholar 

  18. McGookin, R. (1985) RNA Extraction by the Guanidine Thiocyanate Procedure, in Methods in Molecular Biology, Vol. 2 (Walker, J., ed.) Humana, Clifton, New Jersey.

    Google Scholar 

  19. Nelson, C.E. and Ryan, C. A. (1980) In vitro synthesis of pre-proteins of vacuolar compartmented proteinase inhibitors that accumulate in leaves of wounded tomato plants. Proc. Natl. Acad. Sci. USA 77, 1975–1979.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1988 The Humana Press Inc.

About this protocol

Cite this protocol

Slater, A. (1988). Extraction of RNA from Plants. In: Walker, J.M. (eds) New Nucleic Acid Techniques. Methods in Molecular Biology, vol 4. Humana Press. https://doi.org/10.1385/0-89603-127-6:437

Download citation

  • DOI: https://doi.org/10.1385/0-89603-127-6:437

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-127-2

  • Online ISBN: 978-1-59259-491-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics