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Fig. 3 | BMC Cancer

Fig. 3

From: A rational two-step approach to KRAS mutation testing in colorectal cancer using high resolution melting analysis and pyrosequencing

Fig. 3

Outline of the two-step procedure for KRAS codon 12/13 mutation analysis. Genomic DNA from microdissected colorectal cancer cells from FFPE samples is subjected to HRM of a PCR amplicon spanning the mutation-bearing region of interest. For samples that are clearly identified as KRAS mutant or, respectively, WT, the HRM result is incorporated in the final diagnostic report. Samples for which HRM analysis fails technically or which yield ambiguous HRM curves are further evaluated by a second round of HRM and, if results are still invalid, to pyrosequencing. Note that samples for which a WT result is obtained by the diagnostic procedure outlined here require further examination for additional KRAS and NRAS mutations

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