egfr tki response (sensitivity) kit  (Pyrosequencing Inc)

Journal: BMC Cancer

Article Title: Detection and characterization of classical and “uncommon” exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

doi: 10.1186/1471-2407-13-114

Figure Lengend Snippet: 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).

Article Snippet: The mutational analysis was performed by pyrosequencing with PyroMark Q96MA apparatus (Biotage, Uppsala, Sweden) using EGFR TKI response (sensitivity) kit.

Techniques: Mutagenesis

Journal: BMC Cancer

Article Title: Detection and characterization of classical and “uncommon” exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

doi: 10.1186/1471-2407-13-114

Figure Lengend Snippet: Case series characteristics and EGFR mutations distribution

Article Snippet: The mutational analysis was performed by pyrosequencing with PyroMark Q96MA apparatus (Biotage, Uppsala, Sweden) using EGFR TKI response (sensitivity) kit.

Techniques: Biomarker Discovery

Journal: BMC Cancer

Article Title: Detection and characterization of classical and “uncommon” exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

doi: 10.1186/1471-2407-13-114

Figure Lengend Snippet: EGFR exon 19 mutational analysis on 321 prospectively collected samples. The diagram illustrates the results obtained using ARMS technique, EGFR TKI response (sensitivity) kit for pyrosequencing and the home-made dispensation order for EGFR exon 19 mutational analysis. As compared to EGFR TKI response (sensitivity) kit, the NDO allowed to characterize the specific nucleotidic change in the presence of uncommon mutations in a single step, avoiding further need of Sanger sequencing. ( Abbreviations : Pyro-kit: commercial pyrosequencing analysis; Pyro-NDO: novel dispensation order for pyrosequencing; wt : wild type; ARMS: Amplification Refractory Mutation System; mut: mutations).

Article Snippet: The mutational analysis was performed by pyrosequencing with PyroMark Q96MA apparatus (Biotage, Uppsala, Sweden) using EGFR TKI response (sensitivity) kit.

Techniques: Sequencing, Amplification, Mutagenesis

Journal: BMC Cancer

Article Title: Detection and characterization of classical and “uncommon” exon 19 Epidermal Growth Factor Receptor mutations in lung cancer by pyrosequencing

doi: 10.1186/1471-2407-13-114

Figure Lengend Snippet: 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).

Article Snippet: The mutational analysis was performed by pyrosequencing with PyroMark Q96MA apparatus (Biotage, Uppsala, Sweden) using EGFR TKI response (sensitivity) kit.

Techniques: Sequencing, Mutagenesis, Amplification, Comparison, Control