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Mapping Adenines, Guanines, and Pyrimidines in RNA
Nucleic Acids Research (1977)
  • Helen Donis-Keller
  • Allan M. Maxam, Harvard University
  • Walter Gilbert, Harvard University

The positions of adenines, guanines, and pyrimidines can be determined by partial nuclease digestion of a terminally labeled RNA molecule. In urea, at elevated temperatures, RNase T1 generates a pattern reflecting cleavage at guanines while RNase U2 cleaves only at adenine. A limited alkaline hydrolysis provides a continuum of fragments derived from breaks at every phosphodiester bond. The reaction products are electrophoretically fractionated by size in adjacent lanes of a polyacrylamide gel. An autoradiograph of the gel displays the sequence up to 100 nucleotides from the end of the molecule, although uracil cannot as yet be distinguished from cytosine. These techniques form the basis of an RNA sequencing method and are demonstrated on yeast 5.8S ribosomal RNA.

Publication Date
August, 1977
Publisher Statement
© 1977 Oxford University Press. This article was published in Nucleic Acids Research, vol. 4, iss. 8, pages 2527-2538 and may be found here.

Citation Information
Helen Donis-Keller, Allan M. Maxam and Walter Gilbert. "Mapping Adenines, Guanines, and Pyrimidines in RNA" Nucleic Acids Research Vol. 4 Iss. 8 (1977)
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