Advances, challenges, and opportunities in DNA sequencing technology

Nature is composed of an extremely diverse population of organisms with different metabolic abilities that can be mined for tailored applications in the biotechnology and synthetic biology disciplines. Biological information stored within genomes is encoded by a combination of four nucleotides and organized into structural and regulatory units. This information needs to be read and interpreted by specific cellular machineries to generate a functional output. The knowledge of genome sequences from natural origin is essential to dissect and define the information blocks that are the basis of the biological modules defined by the synthetic biology principles. Moreover, the application of synthetic biology for the construction of biological systems with new functions also requires the proper characterization of their working code. Since the invention of chemical and enzymatic DNA sequencing by Maxam, Gilbert, and Sanger, different protocols have evolved during the last 40 years. The establishment of the next-generation sequencing technologies as the routine methodology has increased the universal resources of genetic information by characterizing thousands of different genomes. In this chapter, we analyze the evolution, technical approaches, and methodologies of the different generations of sequencing protocols and their applications in the field of synthetic biology.