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Science 10 June 1988:
Vol. 240. no. 4858, pp. 1520 - 1522
DOI: 10.1126/science.2453926
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Articles
Science, Vol 240, Issue 4858, 1520-1522
Copyright © 1988 by American Association for the Advancement of Science
DNA and RNA sequence determination based on phosphorothioate chemistry
G Gish
and
F Eckstein
Max-Planck-Institut fuer experimentelle Medizin, Abteilung Chemie, Goettingen, West Germany.
The difference in reactivity between phosphate and phosphorothioate diesters is the basis of a chemical degradation scheme for the sequencing of DNA and RNA. The phosphorothioate groups are incorporated into the nucleic acid in four separate enzymatic reactions, with three of the natural nucleoside triphosphates and one alpha-thiotriphosphate in each reaction. Selective strand cleavage is achieved through alkylation to form the hydrolytically labile phosphorothioate triester. As an example, the sequence analysis is presented of M13 phage DNA and of RNA prepared by transcription with SP6 RNA polymerase.
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