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Fig. 2 | BMC Cancer

Fig. 2

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

Fig. 2

Sensitivity of pyrosequencing. Dilutions of genomic DNA from the KRAS mutant cell line PL45 (codon 12 GGT → GAT, heterozygous mutation) in WT genomic DNA was analysed by pyrosequencing on the Pyromark Q24 platform. The WT sequence and the sequence to analyse including wobble bases at the potentially mutant positions in the region of interest (codons 12 and 13) are indicated in the top panel. Lower panels: KRAS codon 12/13 pyrograms for different dilutions of PL45-DNA in WT DNA. Clear mutation calls are obtained for samples containing ≥25 % of PL45-DNA, corresponding to 12.5 % mutant alleles. For lower concentrations, yielding signals below the LOD of the mutation (2.2 %) + 3 %, presence of a potential low level mutation is suggested, which requires technical and/or biological replication of the analysis

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