Pyrosequencing of clinically relevant polymorphisms

Methods Mol Biol. 2005;311:97-114. doi: 10.1385/1-59259-957-5:097.


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 tri-allelic, indel, and short-repeat polymorphisms, as well as determining allele percentages for methylation or pooled sample assessment. Assays details for Pyrosequencing of clinically relevant polymorphisms are described in this chapter.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Alleles
  • Cytochrome P-450 CYP3A
  • Cytochrome P-450 Enzyme System / genetics
  • Genetic Predisposition to Disease
  • Genotype
  • Humans
  • Multidrug Resistance-Associated Proteins / genetics
  • Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide
  • Sequence Analysis, DNA / methods*


  • Multidrug Resistance-Associated Proteins
  • Adenosine Triphosphate
  • Cytochrome P-450 Enzyme System
  • CYP3A protein, human
  • Cytochrome P-450 CYP3A
  • multidrug resistance-associated protein 1