Nucleotide sequences and representative pyrograms of the commercial kit and novel dispensation order for EGFR exon 19 mutation analysis. (A) In the home-made dispensation order a number of nucleotides major than commercial is present to characterize a wider group of deleted sequences (classical and uncommon) than commercial kit. Circles exemplify anomalous nucleotides identified by NDO only. Example of pyrograms obtained using the commercial KDO (left panels) and NDO (right panels) for pyrosequencing in samples with EGFR exon 19 (B) wild type, (C) classical deletion (c.2235-2249del15), (D) uncommon deletion (c.2240-2257del18). Arrows indicate peak modifications corresponding to anomalous nucleotides identified by NDO only (Abbreviations: KDO: kit dispensation order; NDO: novel dispensation order; WT: wild type).