Fluorescent Sequencing for Heterozygote Mutation Detection

  • Colin A. Graham
  • Alison J. M. Hill
Part of the Methods in Molecular Biology™ book series (MIMB, volume 167)


Direct sequencing of PCR products using the dideoxy chain termination procedure developed by Sanger et al. (1) is now the most commonly used method for defining specific mutations. The main benefits of this method lie in its ease of use and this has been enhanced in recent years by the introduction of fluorescent labels and automated detection systems which obviate the need for radioactivity. Although the initial purchase price for automated sequencers is high, this is compensated for by single-tube reaction chemistry and rapid analysis and base calling.


Polymerase Chain Reaction Product Polymerase Chain Reaction Reaction Sequence Profile Cycle Sequencing Reaction 373A Instrument 
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Copyright information

© Humana Press Inc. 2001

Authors and Affiliations

  • Colin A. Graham
    • 1
  • Alison J. M. Hill
    • 2
  1. 1.Northern Ireland Regional Genetics CentreBelfast City Hospital TrustNorthern Ireland, UK
  2. 2.Department of Anatomy and PhysiologyUniversity of DundeeScotland, UK

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