Massively Parallel (“Next-Generation”) DNA Sequencing

Featured Article: Margulies M, Egholm M, Altman WE, Attiya S, Bader JS, Bemben LA, et al. Genome sequencing in microfabricated high-density picolitre reactors. Nature 2005;437:376–80.4 Since Watson and Crick's 1953 landmark discovery that biological information was encoded as a sequence of chemical building blocks (DNA), a significant component of genetic research has been devoted to not only establishing disease–gene associations but also identifying genetic determinants for human quantitative traits such as blood pressure, body mass index, and pigmentation. Essential to this research is the technology to accurately characterize component nucleotides in exactly the order in which they naturally appear: “sequencing” of DNA. The first developed technologies, Maxam-Gilbert's strand cleavage-based techniques (1976) and the Sanger dideoxy chain-termination (1978), sequenced disease-gene candidate regions that were identified through linkage mapping, with subsequent isolation of both the responsible gene and the spectrum of causative mutations. Based on these successes, the Human Genome Project was launched in 1990 and, in 2001, buoyed by Sanger sequencing, the first human reference genome was published (1). Although that article formally marked the transition to the “genomics era,” its cost of $2.7 billion and 10 years of extensive human capital across 20 collaborative centers worldwide highlighted to us that the process of sequencing would need to be reengineered if translating personalized medicine into the clinic for individual patients were to be feasible. In 1999, we founded 454 Life Sciences to address these needs, and in the article featured here, we reported on the 3 fundamental engineering principles that not only drove our first-to-market next-generation DNA sequencing (NGS) platform, but still remain as the anchoring features of all subsequent technologies: monoclonality by limiting dilution, sequencing by synthesis, and the monolithic idea.