Single Nucleotide Polymorphisms and DNA Methylation Analysis Using Pyrosequencing Methods

The data generated from the Human Genome Project has led to an explosion of technology for low-, medium-, and high-throughput genotyping methods. Pyrosequencing is a genotyping assay based on sequencing by synthesis. Short runs of sequence around each polymorphism are generated, allowing for internal controls for each sample. Pyrosequencing can also be used to identify triallelic, indel, and short-repeat polymorphisms, as well as determining allele percentages for DNA methylation or pooled sample assessment. Assays details for pyrosequencing of clinically relevant polymorphisms and DNA methylations are described in this chapter.