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Fig. S1. An example of a calibration curve between inoculated cDNA ratio and ratio of amplified fragments. Horizontal axis (normal epidermal growth factor receptor [EGFR] cDNA)/(L858R EGFR cDNA). Vertical axis (fragment with ROX-labeled T)/(fragment with R110-labeled G), adjusted by relative value of intrinsic fluorescence intensities of both dyes. (Method of construction of the calibration curves.) cDNA fragments derived from the normal and mutated (L858R) EGFR genes were cloned. To mimic the experimental conditions, cloned mutated cDNA was diluted to the amount of genomic DNA purified from a 2.5 × 104-µm2 tissue section, the amount used for a single assay. The amount of genomic DNA was estimated from the copy number of the aldolase gene measured by real-time polymerase chain reaction (PCR). Mixtures (2 : 1, 1 : 1, 1 : 2) of normal and mutated cloned cDNA were subjected to PCR amplification and the SNaP shot assay. The calibration curve represents the average of 10 repeated experiments.

Fig. S2. Kaplan–Meier analysis of patients heterozygous for epidermal growth factor receptor (EGFR) mutation (patients 1–8, dotted line) and those with the dominant EGFR mutation allele (patients 9–15, straight line). (a) Time to disease progression after gefitinib treatment. (b) Overall survival after gefitinib treatment. Vertical axis, fraction of patients with no progression (a) or patient survival (b). Horizontal axis, time in months.

Table S1. Sequences of oligonucleotide primers used for polymerase chain reaction (PCR) amplification of the epidermal growth factor receptor (EGFR) exons. Only exon 19 was amplified with nested PCR. The internal primers are marked by ‘int’

Table S2. Sequences of oligonucleotide primers used for the SNaP shot assay

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