Abstract
Despite the influx of high throughput sequencing techniques, there is still a niche for low-medium throughput genotyping technologies for small-scale screening and validation purposes. 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 utilized to identify tri-allelic, indel, and short repeat polymorphisms, as well as determining allele percentages for methylation or pooled sample assessment. This range of applications makes it well-suited to the research laboratory as a one-stop system.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Evans WE, McLeod HM (2003) Pharmacogenomics—drug disposition, drug targets, and side effects. N Engl J Med 348: 538–549
Marsh S (2009) Pyrosequencing. In: Patrinos GP, Ansorge W (eds) Molecular diagnostics. Elsevier, USA
Freimuth RR, Ameyaw M-M, Pritchard SC, Kwok P-Y, McLeod HL (2004) High-throughput genotyping methods for pharmacogenomic studies. Curr Pharmacogenomics 2:21–33
Ahluwalia R, Freimuth R, McLeod HL, Marsh S (2003) Use of pyrosequencing to detect clinically relevant polymorphisms in dihydropyrimidine dehydrogenase. Clin Chem 49:1661–1664
Hoskins JM, Marcuello E, Altes A, Marsh S, Maxwell T, Van Booven DJ, Pare L, Culverhouse R, McLeod HL, Baiget M (2008) Irinotecan pharmacogenetics: influence of pharmacodynamic genes. Clin Cancer Res 14:1788–1796
Saeki M, Saito Y, Jinno H, Tohkin M, Kurose K, Kaniwa N, Komamura K, Ueno K, Kamakura S, Kitakaze M, Ozawa S, Sawada J (2003) Comprehensive UGT1A1 genotyping in a Japanese population by pyrosequencing. Clin Chem 49:1182–1185
Garsa A, Marsh S, McLeod HL (2005) CYP3A4 and CYP3A5 genotyping by Pyrosequencing. BMC Med Genet 6:19
Rose CM, Marsh S, Ameyaw MM, McLeod HL (2003) Pharmacogenetic analysis of clinically relevant genetic polymorphisms. Methods Mol Med 85:225–237
Lee SS, Kim WY, Jang YJ, Shin JG (2008) Duplex pyrosequencing of the TPMT3C and TPMT6 alleles in Korean and Vietnamese populations. Clin Chim Acta 398:82–85
Yu J, Marsh S (2008) SNP and DNA methylation analysis with Pyrosequencing. In: Wang F (ed) Biomarker methods in drug discovery and development: methods and protocols. Humana, Totowa, pp 119–140
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
King, C.R., Marsh, S. (2013). Pyrosequencing of Clinically Relevant Polymorphisms. In: Innocenti, F., van Schaik, R. (eds) Pharmacogenomics. Methods in Molecular Biology, vol 1015. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-435-7_6
Download citation
DOI: https://doi.org/10.1007/978-1-62703-435-7_6
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-434-0
Online ISBN: 978-1-62703-435-7
eBook Packages: Springer Protocols