A novel “sequencing-by-synthesis” principle for DNA sequencing was developed, taking advantage of the detection of pyrophosphate release through a combination of enzymes to generate light. Pyrosequencing, first described in 1993, was further developed in the United States into the first “next-generation” DNA sequencing instruments (Roche 454 sequencers), starting a new era in genomics research. In the HPA effort, next-generation sequencing has been used for genome-wide transcriptomics profiles of the human protein-coding genes.
- Nyrén P et al., Solid phase DNA minisequencing by an enzymatic luminometric inorganic pyrophosphate detection assay. Anal Biochem. (1993)
PubMed: 8382019 DOI: 10.1006/abio.1993.1024
Other selected publications
Ronaghi M et al., Real-time DNA sequencing using detection of pyrophosphate release. Anal Biochem. (1996)
PubMed: 8923969 DOI: 10.1006/abio.1996.0432
Ronaghi M et al., A sequencing method based on real-time pyrophosphate. Science. (1998)
PubMed: 9705713 DOI: 10.1126/science.281.5375.363
Margulies M et al., Genome sequencing in microfabricated high-density picolitre reactors. Nature. (2005)
PubMed: 16056220 DOI: 10.1038/nature03959
Figure legend: The principle of pyrosequencing involves iterative additions of the four nucleotides and detection of light to monitor incorporation of a nucleotide by a DNA polymerase. Adapted from M. Ronaghi et al. (1998).
- First method for pyrosequencing was published in 1993
- Pyrosequencing opened the new era of “next-generation sequencing,” leading to a rapid lowering of the cost for DNA sequencing
- Used by the HPA consortium for genome-wide transcriptomics analysis
- A search for “pyrosequencing” in Google Scholar yields more than 100,000 publications
- A search for “next-generation sequencing” in Google Scholar yields more than 700,000 publications