Sanger, F.; Coulson, A. R.

Journal of Molecular Biology (1975), 94(3), 441-8 CODEN: JMOBAK; ISSN: 0022-2836. English.

A simple and rapid method was described for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase. The method used 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 2 sequences in bacteriophage .vphi.X174 DNA by using the synthetic decanucleotide A-G-A-A-A-T-A-A-A-A and a restriction enzyme digestion product as primers.