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PDF file - 365K, Supplemental Figure 1: Assay optimization. For each TaqMan probe, the optimal annealing temperature was determined by testing each assay across a temperature gradient of 55.0 - 65{degree sign}C. Typical FAM plots for EGFR L858R (A) and EGFR exon 19 deletion (B) are shown. Supplemental Figure 2: Assay characteristics. As the sample input increases, the copies/μL output increases in a linear fashion across a wide dynamic range for both the L858R assay (A) and the exon 19 deletion assay (B). Testing for 10 and 50 copies of mutant EGFR in a background of 1000 and 50,000 genome equivalents (GE), the L858R assay demonstrates more consistent sensitivity (C) than the exon 19 deletion assay (D). Supplemental Figure 3: Detection of BRAF V600E in cfDNA from patients with advanced melanoma. Supplemental Figure 4: Correlation of LINE-1 quantitative PCR (QPCR) levels with DNA concentration as measured with PicoGreen across 69 plasma specimens (R2 = 0.94, p |
