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High-Resolution Genotyping by Amplicon Melting Analysis Using LCGreen

Clinical Chemistry · 2003 · Vol. 49(6) · pp. 853–860

Carl T. Wittwer✉(University of Utah)G. H. Reed(University of Utah)Cameron N. Gundry(University of Utah)Joshua G Vandersteen(University of Utah)Robert J. Pryor(University of Utah)

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Abstract

High-resolution melting analysis of PCR products amplified in the presence of LCGreen can identify both heterozygous and homozygous sequence variants. The technique requires only the usual unlabeled primers and a generic double-stranded DNA dye added before PCR for amplicon genotyping, and is a promising method for mutation screening.

Molecular Biology Techniques and Applications CRISPR and Genetic Engineering DNA and Nucleic Acid ChemistryAmpliconHigh Resolution MeltMelting curve analysisGenotypingMolecular biologyPrimer (cosmetics)BiologyPolymerase chain reactionOligonucleotidegenomic DNA

MeSH terms

Blood ProteinsDiaminesDNA Mutational AnalysisFluorescent DyesGenotypeHemoglobin CHemoglobin, SickleHumansOrganic ChemicalsQuinolinesReceptors, SerotoninPolymerase Chain ReactionCystic Fibrosis Transmembrane Conductance RegulatorHeteroduplex AnalysisPolymorphism, Single Nucleotide

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