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Open AccessJournal ArticleDOI

DNA sequencing with chain-terminating inhibitors

Frederick Sanger, +2 more
- 01 Dec 1977 - 
- Vol. 74, Iss: 12, pp 5463-5467
TLDR
A new method for determining nucleotide sequences in DNA is described, which makes use of the 2',3'-dideoxy and arabinon nucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase.
Abstract
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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The human immunodeficiency virus type 1-specific protein vpu is required for efficient virus maturation and release.

TL;DR: Indirect immunofluorescence analyses of cultures inoculated with wild-type virus with use of a vpu-specific antiserum demonstrated that vpu is mainly localized to a perinuclear region in the cytoplasm of virus-producing cells.
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KAR3, a kinesin-related gene required for yeast nuclear fusion

TL;DR: The phenotypes of kar3 mutants suggest that the protein mediates microtubule sliding during nuclear fusion and possibly mitosis and a larger family of kinesin-like proteins characterized by the presence of the mechanochemical domain tethered to different protein binding domains.
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Nanopores and nucleic acids: prospects for ultrarapid sequencing

TL;DR: Nanopore detection and characterization of single molecules represents a new method for directly reading information encoded in linear polymers and it is possible that nucleic acid sequences can be determined at rates exceeding a thousand bases per second.
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Plasma and cytoplasmic gelsolins are encoded by a single gene and contain a duplicated actin-binding domain.

TL;DR: Southern blot analysis indicates that a single gene in the haploid genome encodes both protein forms, and the inferred amino-acid sequence reveals the presence of a signal peptide, a long tandem repeat that matches the actin-binding domains of gelsolin, a tetrapeptide present in actin and extended regions of identical sequence with rabbit macrophages.
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Use of degenerate oligonucleotides for amplification of the nifH gene from the marine cyanobacterium Trichodesmium thiebautii.

TL;DR: Examination of the DNA and presumed amino acid sequence shows that the gene is most closely related to that of Anabaena spp.
References
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Journal ArticleDOI

A new method for sequencing DNA

TL;DR: Reactions that cleave DNA preferentially at guanines, at adenines,At cytosines and thymines equally, and at cytosine alone are described.
Journal ArticleDOI

A rapid method for determining sequences in DNA by primed synthesis with DNA polymerase.

TL;DR: A simple and rapid method for determining nucleotide sequences in single-stranded DNA by primed synthesis with DNA polymerase is described and was used to determine two sequences in bacteriophage φX174 DNA.
Journal ArticleDOI

Nucleotide sequence of bacteriophage G4 DNA.

TL;DR: The sequence identifies many of the features responsible for the production of the proteins of the nine known genes of the organism, including initiation and termination sites for the proteins and RNAs.
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