Next-Generation Sequencing

Next-generation sequencing (NGS), otherwise known as deep or massively parallel sequencing, refers to the technological advances in DNA sequencing instrumentation that enable the generation of hundreds of thousands to millions of sequence reads per run. Sequencing of the human genome, which was once a >10-year endeavor by the NIH at the cost of approximately $3 billion (1), can now be done routinely on a single instrument. Rapid advances in technology led to the first-ever FDA clearance of an NGS instrument, the Illumina MiSeq, in 2014 (2), and the development of rapid, miniaturized sequencing devices such as the Oxford Nanopore are ongoing (3). The applications of NGS are wide-ranging and include (i) wholegenome sequencing, (ii) pathogen discovery, (iii) metagenomic/microbiome analyses, (iv) transcriptome profiling, and (vi) infectious disease diagnosis. Here we will focus on NGS technology and the last three applications, because the first two topics are described in detail elsewhere.