Pyrosequencing in cancer

Today, DNA sequencing is one of the most important tools for the study of nucleic acid structure and composition. Traditional DNA sequencing methods are based on dideoxy chain termination technology and are limited in both throughput and cost for most future applications. Recently, pyrosequencing has surfaced as a new sequencing methodology for real-time nucleotide sequencing. It is a nonelectrophoretic DNA sequencing method which circumvents the need for labeled nucleotides, labeled primers, and gel electrophoresis, and offers the advantages of high accuracy, flexibility, parallel processing, and easy automation. Thus, it is increasingly being used in DNA sequencing, genotyping, single nucleotide polymorphism analysis, allele quantification, and whole genome sequencing within the areas of clinical genetics, pharmacogenetics, and microbiology. In this chapter, we will briefly review the key aspects of pyrosequencing technology with a focus on its application in hotspot mutation analysis.