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Proc Natl Acad Sci U S A. Dec 1977; 74(12): 5463–5467.
PMCID: PMC431765

DNA sequencing with chain-terminating inhibitors


A new method for determining nucleotide sequences in DNA is described. It is similar to the “plus and minus” method [Sanger, F. & Coulson, A. R. (1975) J. Mol. Biol. 94, 441-448] but makes use of the 2′,3′-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase. The technique has been applied to the DNA of bacteriophage ϕX174 and is more rapid and more accurate than either the plus or the minus method.

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Selected References

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  • Sanger F, Coulson AR. A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase. J Mol Biol. 1975 May 25;94(3):441–448. [PubMed]
  • Sanger F, Air GM, Barrell BG, Brown NL, Coulson AR, Fiddes CA, Hutchison CA, Slocombe PM, Smith M. Nucleotide sequence of bacteriophage phi X174 DNA. Nature. 1977 Feb 24;265(5596):687–695. [PubMed]
  • Maxam AM, Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. [PMC free article] [PubMed]
  • Atkinson MR, Deutscher MP, Kornberg A, Russell AF, Moffatt JG. Enzymatic synthesis of deoxyribonucleic acid. XXXIV. Termination of chain growth by a 2',3'-dideoxyribonucleotide. Biochemistry. 1969 Dec;8(12):4897–4904. [PubMed]
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  • Geider K. DNA synthesis in nucleotide-permeable Escherichia coli cells. The effects of nucleotide analogues on DNA synthesis. Eur J Biochem. 1972 Jun 9;27(3):554–563. [PubMed]
  • McCarthy JR, Jr, Robins MJ, Townsend LB, Robins RK. Purine nucleosides. XIV. Unsaturated furanosyl adenine nucleosides prepared via base-catalyzed elimination reactions of 2'-deoxyadenosine derivatives. J Am Chem Soc. 1966 Apr 5;88(7):1549–1553. [PubMed]
  • Büchi H, Khorana HG. CV. Total synthesis of the structural gene for an alanine transfer ribonucleic acid from yeast. Chemical synthesis of an icosadeoxyribonucleotide corresponding to the nucleotide sequence 31 to 50. J Mol Biol. 1972 Dec 28;72(2):251–288. [PubMed]
  • Robins MJ, McCarthy JR, Jr, Robins RK. Purine nucleosides. XII. The preparation of 2',3'-dideoxyadenosine, 2',5'-dideoxyadenosine, and 2',3',5'-trideoxyadenosine from 2'-deoxyadenosine. Biochemistry. 1966 Jan;5(1):224–231. [PubMed]
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