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Denaturing HPLC for Mutation Screening

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PCR Mutation Detection Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 688))

Abstract

Denaturing High-Performance Liquid Chromatography (dHPLC) is probably the most versatile and one of the most widely used mutation screening technologies. It benefits from a combination of relative technical simplicity and a very high sensitivity (mutation detection rate), approaching 100%. DHPLC can reliably detect single-base mismatches in fragments between 150 and 500 bp, although detection in fragments up to 1,500 bp has been reported. The ability of dHPLC to detect both known and unknown mutations/SNPs, and its’ high sensitivity and specificity (reproducibility) has put this technology at the forefront of genetic analysis for a wide variety of diseases.

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Acknowledgment

Some Figures are reproduced with kind permission from Transgenomic Inc. (Omaha, USA).

Author information

Authors and Affiliations

  1. Molecular Genetics, Centre for Haemostasis & Thrombosis, St. Thomas’ Hospital, Guy’s & St Thomas’ NHS Foundation Trust, London, UK

    Mike Mitchell & Jacqueline Cutler

Authors
  1. Mike Mitchell
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  2. Jacqueline Cutler
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Corresponding author

Correspondence to Jacqueline Cutler .

Editor information

Editors and Affiliations

  1. NHS Foundation Trust, Department of Haematology, Birmingham Childrens Hospital, Steelhouse Lane, Birmingham, B4 6NH, United Kingdom

    Bimal D.M. Theophilus

  2. Dept. Biosciences, University of Hertfordshire, College Lane, Hatfield, Herts., AL10 9AB, United Kingdom

    Ralph Rapley

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© 2011 Humana Press

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Mitchell, M., Cutler, J. (2011). Denaturing HPLC for Mutation Screening. In: Theophilus, B., Rapley, R. (eds) PCR Mutation Detection Protocols. Methods in Molecular Biology, vol 688. Humana Press. https://doi.org/10.1007/978-1-60761-947-5_3

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  • DOI: https://doi.org/10.1007/978-1-60761-947-5_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-946-8

  • Online ISBN: 978-1-60761-947-5

  • eBook Packages: Springer Protocols

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