Abstract
The first large genome fully sequenced by next-generation sequencing (NGS) was that of a bacteriophage using sequencing by synthesis (SBS) as a paradigm. SBS in NGS is underpinned by ‘reversible-terminator chemistry’. To grow from proof of concept to being both affordable and practical, SBS needed to overcome a series of challenges, each of which required the invention of new chemistries. These included the design and synthesis of unnatural deoxynucleotide triphosphates (dNTPs), engineering a suitable polymerase, a new surface chemistry and an ingenious molecular solution to neutralize copying errors inherent to all polymerases. In this historical Perspective, we discuss how NGS was developed from Sanger sequencing, highlighting the chemistry behind this technology, which has impacted biology in unprecedented ways.
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Acknowledgements
We thank all those whose intellectual and practical contributions have collectively enabled the development of NGS. The intellectual property underlying NGS is protected by a series of patents not discussed in this article. We thank J. Szostak, V. Ramakrishnan and J.-M. Lehn for fruitful discussions and our colleagues at the University of Chicago and the Institut Curie for their constructive comments. R.R. was supported by the European Research Council under the European Union’s Horizon 2020 research and innovation programme (grant agreement no. 647973), Foundation Charles Defforey-Institut de France and Ligue Contre le Cancer. Y.K. is supported by NIH grant 1DP1GM149751-01 and the Ono Pharma Foundation.
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Rodriguez, R., Krishnan, Y. The chemistry of next-generation sequencing. Nat Biotechnol 41, 1709–1715 (2023). https://doi.org/10.1038/s41587-023-01986-3
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DOI: https://doi.org/10.1038/s41587-023-01986-3