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Next generation sequencing (Pyrosequencing)

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 extensively for genome-wide transcriptomics profiles of the human protein-coding genes.

Key publication

  • 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

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).

Key facts

  • 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