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Targeted Next-generation Sequencing and Bioinformatics Pipeline to Evaluate Genetic Determinants of Constitutional Disease
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The chemistry of next-generation sequencing
Raphaël Rodriguez1, Yamuna Krishnan2,3,4
1Institut Curie, CNRS, INSERM, PSL Research University, Equipe Labellisée Ligue Contre le Cancer, Paris, France. raphael.rodriguez@curie.fr.
Nature Biotechnology
|October 16, 2023
Summary
Next-generation sequencing (NGS) revolutionized genomics using reversible-terminator chemistry. Overcoming challenges in unnatural deoxynucleotide triphosphates (dNTPs) and polymerase engineering made SBS sequencing affordable and practical.
Area of Science:
- Genomics
- Molecular Biology
- Biochemistry
Background:
- Next-generation sequencing (NGS) emerged as a powerful tool in genomics.
- The initial large genome sequenced by NGS was a bacteriophage, utilizing sequencing by synthesis (SBS).
- SBS technology relies on 'reversible-terminator chemistry'.
Purpose of the Study:
- To provide a historical perspective on the development of NGS.
- To highlight the crucial role of chemistry in advancing SBS technology.
- To discuss the innovations that enabled NGS to become affordable and practical.
Main Methods:
- Review of historical development from Sanger sequencing to NGS.
- Focus on the chemical innovations underpinning sequencing by synthesis.
- Discussion of challenges and solutions in polymerase and surface chemistry.
Main Results:
- The development of unnatural deoxynucleotide triphosphates (dNTPs) was essential.
- Engineering of suitable polymerases and new surface chemistries were critical.
- Molecular solutions were devised to mitigate polymerase copying errors.
Conclusions:
- NGS, driven by chemical advancements, has profoundly impacted biological research.
- The evolution of SBS chemistry was key to making large-scale genome sequencing accessible.
- Innovations in chemistry were fundamental to the success and widespread adoption of NGS.
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