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Clinical Applications of Capillary Electrophoresis pp 199–210Cite as

Triplet-Repeat Primed PCR and Capillary Electrophoresis for Characterizing the Fragile X Mental Retardation 1 CGG Repeat Hyperexpansions

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1972))

Abstract

Fragile X mental retardation 1 (FMR1) CGG repeat expansions cause fragile X syndrome—the leading monogenic form of intellectual disability—and increase the risk for fragile X-associated tremor ataxia syndrome and fragile X-associated primary ovarian insufficiency. Southern blot (SB) analysis is the current gold standard test for FMR1 molecular diagnosis. Several polymerase chain reaction (PCR)-based methods are now available for sizing FMR1 CGG repeat expansions. These methods offer higher diagnostic sensitivity and specificity compared to SB analysis, significantly reduce the turnaround time and increase throughput. In this chapter, we describe a triplet-repeat primed PCR protocol that employs capillary electrophoresis to resolve the derived amplicon products, enabling precise determination of the FMR1 genotypes in both males and females and characterization of the CGG repeat structure.

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Author information

Authors and Affiliations

  1. Department of Pediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Indhu-Shree Rajan-Babu & Samuel S. Chong

  2. Department of Medical Genetics, The University of British Columbia, Vancouver, BC, Canada

    Indhu-Shree Rajan-Babu

  3. Khoo Teck Puat—National University Children’s Medical Institute, National University Health System, Singapore, Singapore

    Samuel S. Chong

  4. Department of Laboratory Medicine, National University Hospital, Singapore, Singapore

    Samuel S. Chong

Authors
  1. Indhu-Shree Rajan-Babu
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  2. Samuel S. Chong
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Corresponding author

Correspondence to Samuel S. Chong .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutics, School of Pharmacy, Virginia Commonwealth University, Richmond, VA, USA

    Terry M. Phillips

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Rajan-Babu, IS., Chong, S.S. (2019). Triplet-Repeat Primed PCR and Capillary Electrophoresis for Characterizing the Fragile X Mental Retardation 1 CGG Repeat Hyperexpansions. In: Phillips, T.M. (eds) Clinical Applications of Capillary Electrophoresis. Methods in Molecular Biology, vol 1972. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9213-3_14

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  • DOI: https://doi.org/10.1007/978-1-4939-9213-3_14

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9212-6

  • Online ISBN: 978-1-4939-9213-3

  • eBook Packages: Springer Protocols

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