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Nucleic Acids Research, 1978, Vol. 5, No. 9 3129-3140
© 1978

Articles

3' End labelling of RNA with 32P suitable for rapid gel sequencing

G. Winter and G.G. Brownlee

MRC Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, UK

Received July 12, 1978. A new general method of labelling the 2',3'-diol end of RNA with 32P has been devised suitable for gel sequencing. Poly(A) polymerase (E. coli) is incubated with the RNA and limiting amounts of {alpha}-32P-ATP. The mono-addition product iS then cleaved with periodate and ß-eliminated with aniline, leaving the RNA terminally labelled with 3' 32P-phosphate. When applied to a model compound, tRNAPhe from E. coli, over 28 residues could be read from the 3' end.


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