Nucleic Acids pp 125-130 | Cite as

The Estimation of mRNA Content by Poly(U) Hybridization

  • Robert J. Slater
Part of the Methods in Molecular Biology book series (MIMB, volume 2)

Abstract

The sequence of poly(adenylic) acid, present at the 3′ end of the majority of eukaryotic mRNA molecules, forms the basis of a sensitive technique for the estimation of mRNA content in nucleic acid samples. Under suitable conditions, poly(A) will form RNA-RNA hybrids with poly(U) in vitro. The poly (A) content of RNA samples can therefore be detected by hybridization with saturating amounts of 3H-poly(U) (1,2). Following the removal of excess 3H-poly(U) by ribonuclease treatment, the hybrids can be collected by TCA precipitation and quantified by scintillation counting. If the results are compared with data obtained from a parallel experiment using known amounts of poly (A), a value for the poly (A) content of any number of RNA preparations can be obtained. The technique can be used to detect less than 10−10g of poly(A).

Keywords

Citrate Polypropylene 

References

  1. 1.
    Covey, S. N., and Grierson, D. (1976) The measurement of plant polyadenylic acid by hybridization with radioactive polyuridylic acid. Planta 131, 75–79.CrossRefGoogle Scholar
  2. 2.
    Bishop, J. O., Rosbash, M., and Evans, D. (1974) Polynucleotide sequences in eukaryotic DNA and RNA that form ribonuclease-resistant complexes with polyuridylic acid. J. Mol. Biol. 85, 75–86.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc. 1984

Authors and Affiliations

  • Robert J. Slater
    • 1
  1. 1.Division of Biological and Environmental SciencesThe Hatfield PolytechnicHatfieldEngland

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