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Sanger

Sanger sequencing, also known as the "chain termination method," is a DNA sequencing technique that was developed by the English biochemist Frederick Sanger and his colleagues in 1977 1. This method is designed for determining the sequence of nucleotide bases in a piece of DNA (commonly less than 1,000 bp in length) with 99.99% base accuracy 2. These fragments are then used to assemble larger DNA fragments and, eventually, entire chromosomes.

First, feel free to watch this video to get an overview of how Sanger sequencing works.

  1. Sanger, F., Nicklen, S., & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proceedings of the national academy of sciences, 74(12), 5463-5467. doi: 10.1073/pnas.74.12.5463 ↩

  2. Shendure, J., & Ji, H. (2008). Next-generation DNA sequencing. Nature biotechnology, 26(10), 1135-1145. doi: 10.1038/nbt1486 ↩

  3. Al-Shuhaib, M. B. S., & Hashim, H. O. (2023). Mastering DNA chromatogram analysis in Sanger sequencing for reliable clinical analysis. J. Genet. Eng. Biotechnol., 21(1), 115. doi: 10.1186/s43141-023-00587-6 ↩

  4. Arteche-López, A., Ávila-Fernández, A., Romero, R., Riveiro-Álvarez, R., López-Martínez, M. A., Giménez-Pardo, A., ... & Ayuso, C. (2021). Sanger sequencing is no longer always necessary based on a single-center validation of 1109 NGS variants in 825 clinical exomes. Scientific reports, 11(1), 5697. doi: 10.1038/s41598-021-85182-w ↩

  5. De Cario, R., Kura, A., Suraci, S., Magi, A., Volta, A., Marcucci, R., ... & Sticchi, E. (2020). Sanger validation of high-throughput sequencing in genetic diagnosis: Still the best practice?. Frontiers in genetics, 11, 592588. doi: 10.3389/fgene.2020.592588 ↩

  6. Giani, A. M., Gallo, G. R., Gianfranceschi, L., & Formenti, G. (2020). Long walk to genomics: History and current approaches to genome sequencing and assembly. Computational and Structural Biotechnology Journal, 18, 9-19. doi: 10.1016/j.csbj.2019.11.002 ↩

  7. Crossley, B. M., Bai, J., Glaser, A., Maes, R., Porter, E., Killian, M. L., ... & Toohey-Kurth, K. (2020). Guidelines for Sanger sequencing and molecular assay monitoring. Journal of Veterinary Diagnostic Investigation, 32(6), 767-775. doi: 10.1177/1040638720905833 ↩