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Sensitive and Specific KRAS Somatic Mutation Analysis on Whole-Genome Amplified DNA from Archival Tissues  Ronald van Eijk, Marjo van Puijenbroek, Amiet.

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Presentation on theme: "Sensitive and Specific KRAS Somatic Mutation Analysis on Whole-Genome Amplified DNA from Archival Tissues  Ronald van Eijk, Marjo van Puijenbroek, Amiet."— Presentation transcript:

1 Sensitive and Specific KRAS Somatic Mutation Analysis on Whole-Genome Amplified DNA from Archival Tissues  Ronald van Eijk, Marjo van Puijenbroek, Amiet R. Chhatta, Nisha Gupta, Rolf H.A.M. Vossen, Esther H. Lips, Anne-Marie Cleton-Jansen, Hans Morreau, Tom van Wezel  The Journal of Molecular Diagnostics  Volume 12, Issue 1, Pages (January 2010) DOI: /jmoldx Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

2 Figure 1 HRM curves in samples treated with or without WGA. HRM shifted melting curves and difference curves of three different KRAS codon 12 and 13 mutations in duplicate, with or without WGA treatment. g.35G>A (p.12G>D) DNA lower green, wgaDNA upper green; g.38G>A (p.13G>D) DNA lower blue, wgaDNA upper blue; g.35G>T (p.12G>V) red; and a series of eight wild-type samples (gray). The Journal of Molecular Diagnostics  , 27-34DOI: ( /jmoldx ) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

3 Figure 2 Detection limit of KRAS mutations in a PCR of serial diluted wgaDNA. Left panel: Twofold serial dilution of gDNA starting with 10 ng (upper curve) to 0.08 ng of input (lowest curve) shows the detection limit for KRAS mutations in a PCR on wgaDNA. Upper right panel: Sanger sequence on wgaDNA corresponds with 10 ng of gDNA input in the WGA. Lower right panel: Sanger sequence on wgaDNA corresponds with ng of (±100 cells) gDNA input in the WGA. The Journal of Molecular Diagnostics  , 27-34DOI: ( /jmoldx ) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

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