DNA sequencing with chain-terminating inhibitors
F. Sanger, S. Nicklen, A. R. Coulson
Summary
Sanger, Nicklen, and Coulson introduced a new method for determining nucleotide sequences in DNA, building on their earlier 'plus and minus' technique. The method uses 2′,3′-dideoxy and arabinonucleoside analogues of the normal deoxynucleoside triphosphates, which act as specific chain-terminating inhibitors of DNA polymerase, generating a set of partially extended chains that can be size-separated by gel electrophoresis to read the sequence. Applied to bacteriophage φX174 DNA, the approach proved faster and more accurate than the original plus or minus method.
Key findings
- Introduced the use of dideoxynucleoside triphosphate analogues as specific chain-terminating inhibitors of DNA polymerase—the basis of 'dideoxy' (Sanger) sequencing.
- Demonstrated that primer-directed synthesis with chain terminators produces nested DNA fragments that, when separated by gel electrophoresis, reveal the nucleotide sequence.
- Validated the method on bacteriophage φX174 DNA, showing it to be faster and more accurate than the earlier plus-and-minus technique.
Subjects & keywords
Cite this paper
F. Sanger, S. Nicklen, & A. R. Coulson (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences. https://doi.org/10.1073/pnas.74.12.5463
@article{sanger1977sequencing,
author = {F. Sanger and S. Nicklen and A. R. Coulson},
title = {DNA sequencing with chain-terminating inhibitors},
journal = {Proceedings of the National Academy of Sciences},
year = {1977},
doi = {10.1073/pnas.74.12.5463},
url = {https://doi.org/10.1073/pnas.74.12.5463}
}