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Published on: December 2, 2009
The estimation of mRNA content by poly(u) hybridization
1Division of Biological and Environmental Sciences, The Hatfield Polytechnic, Hatfield, Hertfordshire, England.
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 (3)H-poly(U) (1,2). Following the removal of excess (3)H-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(-10)g of poly(A).
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