A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase |
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Authors: | F Sanger AR Coulson |
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Institution: | Medical Research Council Laboratory of Molecular Biology Hills Road, Cambridge CB2 2QH, England |
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Abstract: | A simple and rapid method for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase is described. It depends on the use of Escherichia coli DNA polymerase I and DNA polymerase from bacteriophage T4 under conditions of different limiting nucleoside triphosphates and concurrent fractionation of the products according to size by ionophoresis on acrylamide gels. The method was used to determine two sequences in bacteriophage φX174 DNA using the synthetic decanucleotide A-G-A-A-A-T-A-A-A-A and a restriction enzyme digestion product as primers. |
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