Mutation detection using fluorescent hybridization probes and melting curve analysis.
The LightCycler is a real-time PCR instrument that combines a thermocycler and a micro-volume fluorimeter. LightCycler technology is gaining popularity due to its ability to detect mutations quickly and accurately. Multiple base alterations are discriminated using hybridization probes and fluorescent melting curves. This review focuses on mutation detection and base discrimination by fluorescent hybridization probes. Assay designs for single base mutation detection and complex multiplex reactions are discussed. Types of mutations detected and reported applications are reviewed. Guidelines using melting curve analysis for the clinical laboratory are presented.
Lyon
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Department of Pathology, University of Utah, 50 North Medical Drive, Salt Lake City, UT 84132, USA. lyone@aruplab.com
D004252:DNA Mutational Analysis; D005165:Factor V / Q000235:genetics; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009154:Mutation; D016133:Polymerase Chain Reaction; D013696:Temperature
D005456:Fluorescent Dyes; D005165:Factor V