Figure 3

Sequence analysis of the tumor sample harbouring the novel mutation in EGFR exon 19. (A) ARMS analysis: the amplification of EGFR exon 19 is positive in the cases harbouring the two classical deletions (c.2235-2249del15 and c.2236-2250del15) in comparison to negative wt sample, no template control and the test tumor sample harbouring the new mutation. (B). EGFR TKI response (sensitivity) kit sequencing using commercial dispensation order: the pyrogram is altered but not referable to any classical or uncommon mutation. (C). EGFR TKI response (sensitivity) kit sequencing using NDO: the pyrogram allows to determine a suspected substitution (arrow) of a new nucleotide. (D) Pyrogram trace of the sample obtained using a specific nucleotide dispensation order allows to characterize the new mutation as an insertion of T nucleotide (arrow) in the place of a G nucleotide. (E) Sanger confirmation of the new mutation identified (arrow).