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HyBeacon probes have been used to characterise SNPs in the CYP2C9 and VKORC1 genes associated with variations in the efficiency of warfarin metabolism. PCR amplification of genomic DNA and probe target melting analysis provided a robust and reliable method to differentiate polymorphic sequences at these loci. Probes capped with 5'-trimethoxystilbene were found to exhibit larger fluorescence differences between hybridised and dissociated states than uncapped probes, generating melting peaks of considerably improved height. 3'-Pyrene modifications enhanced probe signal-to-noise and melting peak heights with the additional benefit of acting as PCR stoppers, and 5',3'-doubly capped probes gave the best analytical results.

Original publication

DOI

10.1039/c001177k

Type

Journal article

Journal

Org Biomol Chem

Publication Date

21/06/2010

Volume

8

Pages

2728 - 2734

Keywords

Aryl Hydrocarbon Hydroxylases, Cytochrome P-450 CYP2C9, DNA, Fluorescent Dyes, Genotype, Humans, Mixed Function Oxygenases, Oligonucleotide Probes, Polymerase Chain Reaction, Polymorphism, Genetic, Vitamin K Epoxide Reductases, Warfarin