Mechanisms of Next-Generation Sequencing (NGS)

Abstract

DNA sequencing consists of a number of methodologies, each adopts a unique process of sequencing mechanisms. During 1970s, Sanger sequencing survived the competition against other approaches and dominated DNA sequencing for a number of decades. As stimulated by urgent demand of high throughput sequencing approaches by the Human Genome Project and various genome projects that followed, Next- Generation Sequencing evolved to replace Sanger sequencing as the main sequencing approach. NGS consists of three major sequencing platforms (i.e., 454/Roche, Solexa/Illumina and SOLiD/Life Technologies) and each has its own sequencing mechanism. These mechanisms have experienced severe competition, leading to the election of Illumina system by the sequencing market as the main stream sequencing platform. Before we can fully appreciate the reasons leading to the success of the Illumina system, here we analyze and discuss the sequencing mechanisms adopted by these NGS platforms.

Keywords: 454, Bridge amplification, Emulsion PCR, in situ PCR, Illumina sequencing, Next-generation sequencing, NGS, Solexa, SOLiD, Sequencing-by-synthesis, Sequencing-by-ligation.