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The development and impact of 454 sequencing

Abstract

The 454 Sequencer has dramatically increased the volume of sequencing conducted by the scientific community and expanded the range of problems that can be addressed by the direct readouts of DNA sequence. Key breakthroughs in the development of the 454 sequencing platform included higher throughput, simplified all in vitro sample preparation and the miniaturization of sequencing chemistries, enabling massively parallel sequencing reactions to be carried out at a scale and cost not previously possible. Together with other recently released next-generation technologies, the 454 platform has started to democratize sequencing, providing individual laboratories with access to capacities that rival those previously found only at a handful of large sequencing centers. Over the past 18 months, 454 sequencing has led to a better understanding of the structure of the human genome, allowed the first non-Sanger sequence of an individual human and opened up new approaches to identify small RNAs. To make next-generation technologies more widely accessible, they must become easier to use and less costly. In the longer term, the principles established by 454 sequencing might reduce cost further, potentially enabling personalized genomics.

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Figure 1: Diagram of the pyrosequencing process.
Figure 2: Overview of the 454 sequencing technology.
Figure 3: Next-generation market segmentation.

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Correspondence to Jonathan M Rothberg.

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The authors have financial interests in a number of companies operating in fields that directly benefit from the promotion and success of next-generation sequencing technology in general. Financial interest includes stock ownership, stock options, salaries, consulting fees and bonuses. Currently these financial interests are in RainDance Technologies, and ION Torrent systems. In the past the authors were also key employees, and/or founders, of 454 Life Sciences, are named inventors on key 454 patents, and benefitted from the commercial, scientific and financial success of 454 Life Sciences.

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Rothberg, J., Leamon, J. The development and impact of 454 sequencing. Nat Biotechnol 26, 1117–1124 (2008). https://doi.org/10.1038/nbt1485

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