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Measurement of NOS mRNA by Northern Blotting and the Ribonuclease-Protection Assay

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Nitric Oxide Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 100))

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Abstract

The amount of RNA varies greatly between different cell types and organisms, and the majority of RNA (more than 95%) consists of ribosomal RNA (28S, 18S, and 5S species) and the tRNA. Only a small proportion (usually less than 1% of the total amount) of RNA corresponds to the mRNA species. However, in order to improve mRNA-detection methods, preparations of enriched-mRNA content (poly-A+) can be obtained after elution from oligo-dT columns of total-RNA samples.

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References

  1. Chirgwin, J. M., Przybyla, A. E., MacDonald, R. J., and Rutter, W. J. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18, 5294–5299.

    Article  PubMed  CAS  Google Scholar 

  2. Chomczynski, P. and Sacchi, N. (1987) Single-step method of RNA isolation by acid guanidium thiocyante-phenol-chloroform extraction. Anal. Biochem. 162, 156–159.

    Article  PubMed  CAS  Google Scholar 

  3. Xie, Q-w., Cho, H. J., Calaycay, J., Mumford, R. A., Swiderek, K. M., Lee, T. D., Ding, A., Troso, T., and Nathan, C. F. (1992) Cloning and characterization of inducible nitric oxide synthase from mouse macrophages. Science 256, 225–228.

    Article  PubMed  CAS  Google Scholar 

  4. Gilman, M. (1987) Ribonuclease protection assay, in Current Protocols in Molecular Biology (Ausbel, F. M., Brendt, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds.), Wiley, New York, pp. 4.7.1–4.7.8.

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Author information

Authors and Affiliations

  1. Instituto de Bioquímica CSIC-UCM, Facultad de Farmacia, Universidad Complutense, Madrid, Spain

    Paloma Martín-Sanz

  2. Facultad de Farmacia, Instituto de Bioquímica CSIC-UCM, Universidad Complutense, Madrid, Spain

    Lisardo Boscá

Authors
  1. Paloma Martín-Sanz
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  2. Lisardo Boscá
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Editor information

Editors and Affiliations

  1. University of Sussex, Brighton, UK

    Michael A. Titheradge

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© 1998 Humana Press Inc.

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Martín-Sanz, P., Boscá, L. (1998). Measurement of NOS mRNA by Northern Blotting and the Ribonuclease-Protection Assay. In: Titheradge, M.A. (eds) Nitric Oxide Protocols. Methods in Molecular Biology™, vol 100. Humana Press. https://doi.org/10.1385/1-59259-749-1:163

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  • DOI: https://doi.org/10.1385/1-59259-749-1:163

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-470-9

  • Online ISBN: 978-1-59259-749-9

  • eBook Packages: Springer Protocols

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