KRAS Genotyping of Paraffin-Embedded Colorectal Cancer Tissue in Routine Diagnostics Wilko Weichert, Christiane Schewe, Annika Lehmann, Christine Sers,

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KRAS Genotyping of Paraffin-Embedded Colorectal Cancer Tissue in Routine Diagnostics Wilko Weichert, Christiane Schewe, Annika Lehmann, Christine Sers, Carsten Denkert, Jan Budczies, Albrecht Stenzinger, Hans Joos, Olfert Landt, Volker Heiser, Christoph Röcken, Manfred Dietel The Journal of Molecular Diagnostics Volume 12, Issue 1, Pages 35-42 (January 2010) DOI: 10.2353/jmoldx.2010.090079 Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Different detection methods for KRAS mutations in codon 12/13. A and B: Sanger sequencing. Example of a wild-type (A) and a mutated (B) case (p.G12D). C and D: Pyrosequencing. Example of a wild-type (C) and a mutated (D) case (p.G12D). E–H: Array analysis. Examples of a wild-type case (E) and cases harboring a p.G13D (F), a p.G12D (G), and a p.G12C (H) mutation are presented. I and J: Melting curve analysis. Examples of a wild-type (I) and a mutated (J) case (p.G13D). Arrows indicate location of each mutation. The Journal of Molecular Diagnostics 2010 12, 35-42DOI: (10.2353/jmoldx.2010.090079) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Percentage of mismatches for all detection methods. The percentage was calculated as the number of cases with discordant results (when compared with the other three assays) divided by the total number of tests performed per method. The Journal of Molecular Diagnostics 2010 12, 35-42DOI: (10.2353/jmoldx.2010.090079) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Tissue characteristics and frequency of mutation detection as determined by Sanger sequencing (n = 260). The Journal of Molecular Diagnostics 2010 12, 35-42DOI: (10.2353/jmoldx.2010.090079) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions