Adaptation of an amplicon-based human cancer next-generation sequencing panel assay for murine tumors

• Published in: Analytical biochemistry Vol. 551; pp. 26 - 28
• Main Authors: Meier, David R., Lofgren, Kristopher A., Gurda, Grzegorz T., Kenny, Paraic A.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-06-2018
• Subjects: Animals; Class I Phosphatidylinositol 3-Kinases - genetics; DNA Mutational Analysis - methods; DNA, Neoplasm - genetics; Female; Genes, ras; High-Throughput Nucleotide Sequencing - methods; Humans; Mammary Neoplasms, Experimental - genetics; Mice; Mice, Transgenic; Nucleic Acid Amplification Techniques - methods; Oncogenes; Polymerase Chain Reaction - methods; Protein-Serine-Threonine Kinases - genetics; Species Specificity
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2018.05.007

Unlike humans, inbred genetically engineered mice have minimal inter-individual variation and, consequently, offer substantially increased statistical power for robust definition of recurrent cooperating cancer mutations. While technically feasible, whole exome sequencing is expensive and extremely data-intensive. Somatic mutation analysis using panels of 25–75 genes now provides detailed insight into the biology of human tumors. Here we report an adaptation for mouse tumors of a human PCR amplicon-based panel (Ion Torrent Cancer Hotspot Panel v2) allowing analysis of 18 cancer genes, including Kras, Nras, Hras, Pten, Pik3ca and Smad4, and encompassing regions homologous to more than 2000 known human cancer mutations.