1. Evaluation of BRAF Mutation Testing Methodologies in Formalin-Fixed, Paraffin- Embedded Cutaneous Melanomas 
Johanne Lade-Keller, Kirsten M. Rømer, Per Guldberg, Rikke Riber-Hansen, Lise Lotte Hansen, Torben Steiniche, Henrik Hager, Lasse S. Kristensen 
The Journal of Molecular Diagnostics 
Volume 15, Issue 1, Pages (January 2013)
DOI: /j.jmoldx
Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

2. Figure 1
Selected CADMA, pyrosequencing, and Sanger sequencing results from testing of the cell line dilutions. A: Melting curves from the CADMA experiments. Not all dilutions are shown for clarity. Dilutions containing a theoretical fraction of 50% mutant alleles are represented by the red curves, 30% mutant alleles by the blue curves, 2.5% mutant alleles by the orange curves, 1.25% mutant alleles by the pink curves, % mutant alleles by the green curves, % mutant alleles by the purple curves, and % mutant alleles by the turquoise curves. The 10 wild-type replicates are represented by the brown curves. B: Pyrosequencing results for the dilutions containing a theoretical fraction of 50% and 10% mutant alleles. C: Sanger sequencing results for the dilutions containing a theoretical fraction of 50% and 30% mutant alleles. The position of mutation is indicated by the arrows.
The Journal of Molecular Diagnostics  , 70-80DOI: ( /j.jmoldx )
Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

3. Figure 2
Selected CADMA results from testing of the FFPE malignant melanoma samples. It can be observed from the shape of the melting curves that CADMA can distinguish low-level mutations from high-level mutations. Sample 19 is represented by the light blue melting curves. This sample contained a high fraction of mutated alleles as indicated by the low ΔCt value (2.0) in the TaqMan assay. Sample 24 is represented by the green melting curves. This sample contained a low fraction of mutated alleles as indicated by the high ΔCt value (6.3) in the TaqMan assay. Sample 23 was mutation negative and is represented by the pink melting curves. The brown melting curves represent two wild-type replicates.
The Journal of Molecular Diagnostics  , 70-80DOI: ( /j.jmoldx )
Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

4. Figure 3
Detection of the c.1798_1799GT>AA (V600K) mutation using the CADMA assay. Cell line DNA carrying the c.1798_1799GT>AA mutation was diluted in wild-type DNA to the following fractions: 100% (green curves), 50% (turquoise curve), 25% (orange curve), and 12.5% cell line DNA (pink curve), and compared to melting curves from wild-type samples (brown curve) and c.1799T>A mutated samples (blue curve). It can be observed that the c.1798_1799GT>AA mutation could be distinguished from the wild type in five of five replicates containing 100% cell line DNA. The mutation was not detected in any of the replicates containing lower fractions of cell line DNA. With the exception of the sample containing 100% cell line DNA, only one replicate of each sample is shown for clarity.
The Journal of Molecular Diagnostics  , 70-80DOI: ( /j.jmoldx )
Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions