A new method of DNA sequencing using deoxynucleoside alpha-thiotriphosphates.
A new procedure for determining DNA nucleotide sequences is reported. In the first step of the method, four DNAs, each separately substituted with a different deoxynucleoside phosphorothioate in place of the corresponding monophosphate, are prepared by template-directed polymerization catalyzed by DNA polymerase. In the second step, these DNAs are subjected to stringent exonuclease III treatment, which produces only fragments terminating with a phosphorothioate internucleotide linkage. These can then be separated by standard gel electrophoresis techniques and the sequence can be read directly as in presently used sequencing methods.
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D001483:Base Sequence; D004247:DNA / Q000032:analysis*; D003854:Deoxyribonucleotides; D004591:Electrophoresis, Polyacrylamide Gel; D005090:Exodeoxyribonucleases; D009690:Nucleic Acid Conformation; D013698:Templates, Genetic; D013873:Thionucleotides
D003854:Deoxyribonucleotides; D013873:Thionucleotides; D004247:DNA; D005090:Exodeoxyribonucleases; C020133:exodeoxyribonuclease III