Chapter 8 - DNA sequencing and other methods of exonic and genomic analyses

Abstract

DNA sequencing technologies, other methods of exonic and genomic analysis, and the suitable application of each method are reviewed. The methods described are (1) the Maxam–Gilbert method; (2) the Sanger method; (3) next-generation sequencing (NGS) technologies; (4) methods of exonic and genomic analyses, including fluorescence in situ hybridization analysis, microarray-based array comparative genomic hybridization analysis, polymerase chain reaction (PCR)-based methods [quantitative PCR analysis, multiplex ligation-dependent probe amplification analysis, PCR fragment analysis, and repeat-primed PCR analysis], and Southern blot hybridization analysis. In particular, NGS technologies are currently revolutionizing the field of genomic research, enabling diverse and flexible sequencing applications of genomes, transcriptomes, and epigenomes for various purposes. Genome-wide screening employing NGS technologies demonstrated excellent competence to not only identify causative genes for Mendelian-trait diseases but also identify causative mutations in the context of “clinical sequencing.”