Abstract
Denaturing high-performance liquid chromatography (DHPLC) is an accurate and efficient screening technique used for detecting deoxyribonucleic acid sequence changes by heteroduplex analysis. It can also be used for genotyping of single-nucleotide polymorphisms. The high-sensitivity of DHPLC has made this technique one of the most reliable approaches to mutation analysis and is used in various areas of genetics, both in the research and clinical arena. This chapter describes the methods used for mutation detection analysis and the genotyping of single-nucleotide polymorphisms by DHPLC on the WAVE™ system from Transgenomic Inc.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Oefner, P. J. and Underhill, P. A. (1995) Comparative DNA sequencing by denaturing high-performance liquid chromatography (DHPLC), in ASHG Annual meeting A2666, University of Chicago Press, Chicago.
Navigator Software Manual, Version 1.5.3, © (2003) Transgenomic, Inc., used with permission.
Cotton, R. G. (1997) Slowly but surely towards better scanning for mutations. Trends Genet. 13, 43–46.
O’Donovan, M. C., Oefner, P. J., Roberts, S. C., et al. (1998) Blind analysis of denaturing high-performance liquid chromatography as a tool for mutation detection. Genomics 52, 44–49.
Frueh, F. W. and Noyer-Weidner, M. (2003) The use of denaturing high performance liquid chromatography (DHPLC) for the analysis of genetic variations: impact for diagnostics and pharmacogenetics. Clin. Chem. Lab. Med. 41, 452–461.
Oefner, P. J. (2000) Allelic Discrimination by denaturing high-performance liquid chromatography. J. Chromatogr. B. 739, 345–355.
Devaney, J. M., Pettit, E. L., Kaler, S. G., Vallone, P. M., Butler, J. M., and Marino, M. A. (2001) Genotyping of two mutations in the HFE gene using singlebase extension and high-performance liquid chromatography. Anal. Chem. 73, 620–624.
Wu, G., Hua, L., Zhu, J., Mo, Q., and Xu, X. (2003) Rapid, accurate genotyping of β-thalassaemia mutations using a novel multiplex primer extension/denaturing high-performance liquid chromatography assay. British J. Hematol. 122, 311–316.
Xiao, W. and Oefner, P. J. (2001) Denaturing high-performance liquid chromatography: a review. Human Mutation. 17, 439–474.
Taylor, P., Munson, K., and Gjerde, D. (1999) Detection of mutations and polymorphisms on the WAVE™ DNA Fragment Analysis System. Application Note 101. Transgenomic, Inc.
Kuklin, A., Munson, K., Gjerde, D., Haefele, R., and Taylor, P. (1997/98) Detection of single-nucleotide polymorphisms with the WAVE™ DNA Fragment Analysis System. Genetic Testing 1, 201–206.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Fackenthal, D.L., Chen, P.X., Das, S. (2005). Denaturing High-Performance Liquid Chromatography for Mutation Detection and Genotyping. In: Innocenti, F. (eds) Pharmacogenomics. Methods in Molecular Biology™, vol 311. Humana Press. https://doi.org/10.1385/1-59259-957-5:073
Download citation
DOI: https://doi.org/10.1385/1-59259-957-5:073
Publisher Name: Humana Press
Print ISBN: 978-1-58829-440-1
Online ISBN: 978-1-59259-957-8
eBook Packages: Springer Protocols