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A Multi-Site Study for Detection of the Factor V (Leiden) Mutation from Genomic DNA Using a Homogeneous Invader Microtiter Plate Fluorescence Resonance.

Published byAlvin Gray Modified over 6 years ago

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Presentation on theme: "A Multi-Site Study for Detection of the Factor V (Leiden) Mutation from Genomic DNA Using a Homogeneous Invader Microtiter Plate Fluorescence Resonance."— Presentation transcript:

1 A Multi-Site Study for Detection of the Factor V (Leiden) Mutation from Genomic DNA Using a Homogeneous Invader Microtiter Plate Fluorescence Resonance Energy Transfer (FRET) Assay  Marlies Ledford, Kenneth D. Friedman, Martin J. Hessner, Cynthia Moehlenkamp, Thomas M. Williams, Richard S. Larson  The Journal of Molecular Diagnostics  Volume 2, Issue 2, Pages (May 2000) DOI: /S (10)60623-X Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 Schematic outline of the Invader assay for FVL, showing discrimination of single basepair changes by the Invader assay. The key to single-base discrimination for Invader technology depends both on a match between the WT or Mut probe and the target sequence at the base to be discriminated and an overlap of at least one base by the Invader oligonucleotide. The specific sequence in the target molecule (sample DNA) determines whether cleavage and subsequent fluorescent signal generation will occur. The structures formed in A are substrates recognized by the Cleavase enzyme, whereas the structure shown in B is a structure not recognized by the Cleavase enzyme. The specific base shown in the target sequence is the base to be discriminated. The Journal of Molecular Diagnostics 2000 2, DOI: ( /S (10)60623-X) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Distribution of ratios from the Invader assay results at the study sites for wild-type, heterozygous, and mutant genotypes, as determined by PCR-RFLP or AS-PCR. Note that the scale for the ordinate is logarithmic. The Journal of Molecular Diagnostics 2000 2, DOI: ( /S (10)60623-X) Copyright © 2000 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

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