AS QIAamp DNA FFPE tissue kit (Qiagen, Hilden, Germany) was used to extract DNA from paraffin-embedded tissue samples, and the presence of tumor cells (>70%) was obtained by trimming the normal and necrotic tissues. Genetic analyses of EGFR, KRAS, and BRAF were performed using the OncoCarta Panel14 (link) (version 1.0; Sequenom, San Diego, CA, USA). Mutation data were analyzed using MassArray Typer software version 4.0 (Sequenom). MET skipping mutation was detected by direct sequencing6 (link) using a PTC-200PCR (Bio-Rad, Hercules, CA, USA) with the forward primer 5’-CTTTGTACGTCTCATGTTAT-3’ and reverse primer 5’-CTCCTAGCGACCTAAC-3’. PCR products were purified and labeled using a BigDye Terminator 3.1 cycle-sequencing kit (Applied Biosystems, Foster City, CA), followed by sequencing in an ABI 3500XL Genetic Analyzer (Applied Biosystems).15 (link)
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Genetic Profiling of Tumor Samples
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Corresponding Organization :
Other organizations : Sun Yat-sen University, Sun Yat-sen University Cancer Center
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Variable analysis
independent variables
- DNA extraction using the QIAamp DNA FFPE tissue kit
- Trimming of normal and necrotic tissues to obtain >70% tumor cells
- Genetic analyses of EGFR, KRAS, and BRAF using the OncoCarta Panel14
- Detection of MET skipping mutation by direct sequencing
- Presence of tumor cells (>70%)
- Mutation status of EGFR, KRAS, and BRAF
- Detection of MET skipping mutation
- Paraffin-embedded tissue samples
- No positive or negative controls were explicitly mentioned.
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