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DNA sequencing with chain-terminating inhibitors

Frederick Sanger, +2 more
- 01 Dec 1977 - 
- Vol. 74, Iss: 12, pp 5463-5467
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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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Citations
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The use of double mutants to detect structural changes in the active site of the tyrosyl-tRNA synthetase (Bacillus stearothermophilus)

TL;DR: By making mutants of the Pro51 enzyme at two residues that make hydrogen bonds to the ATP substrate, it is shown that Pro51 greatly improves the strength of one of these contacts.
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mec-3, a homeobox-containing gene that specifies differentiation of the touch receptor neurons in C. elegans

TL;DR: The mec-3 gene is cloned by transposon tagging and it is shown that a meC-3 mutant can be rescued by germ line transformation using a 5.6 kb genomic DNA fragment.
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Gene organization deduced from the complete sequence of liverwort Marchantia polymorpha mitochondrial DNA. A primitive form of plant mitochondrial genome

TL;DR: In this article, the complete sequence of the liverwort mitochondrial DNA was determined and 94 possible genes in the sequence of 186,608 base-pairs were detected, including genes for three species of ribosomal RNA, 29 genes for 27 species of transfer RNA and 30 open reading frames (ORFs) for functionally known proteins.
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Tissue specificity of Drosophila P element transposition is regulated at the level of mRNA splicing

TL;DR: In vitro mutagenesis and genetic analysis demonstrate the existence of a third intron whose removal is required for transposase production and propose that this intron is only removed in the germline and that its removal is the sole basis for the Germline restriction of P element transposition.
Journal ArticleDOI

Two novel GABAA receptor subunits exist in distinct neuronal subpopulations

TL;DR: Cellular localization of the mRNAs encoding the gamma 2 and delta subunits in rat brain revealed that largely distinct neuronal subpopulations express the two subunits, implications for the composition of two different GABAA receptor subtypes and for information processing in networks using GABA for signaling.
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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