An overview of DNA analytical methods

Abstract

The development of rapid parallel sequencing in the last 20 years has begun a revolution in the field of genetics that is changing nearly all disciplines within biology and medicine. Genomic sequencing has become crucial to the diagnosis and clinical management of patients with constitutional diseases and cancer and has quickly become an integral part of the new era of personalized and precision medicine. The precision medicine initiative, released by the NIH in 2015, has catapulted genomic technologies to the forefront of the practice of medicine and biomedical research. This chapter focuses on the core technologies driving the genomic revolution from first generation (Sanger) sequencing to microarray-based technologies, to second, commonly referred to as next-generation sequencing (NGS) methods, and finally to the emerging third generation technologies capable of performing single-molecule and long-read sequencing. The goal of the chapter is to provide a broad overview of these methods of DNA analysis and highlight their strengths and weaknesses. Furthermore, with a knowledge of the different mutation types, we seek to provide the basis for understanding how these technologies work, and can be adopted, to explore other type of nucleic acids and epigenetic changes.