NSCLC Cell Line Characterization

NSCLC cell lines with EML4-ALK [27 (link)], H2228 (CRL-5935, ATCC, LCG standards, Wesel, Germany), and H3122 (Adi F. Gazdar, UT Southwestern, Dallas, TX, USA), or without EML4-ALK, HCC827 (CRL-2868, ATCC, LCG standards, Wesel, Germany), and A549 (CCL-185, ATCC, LCG standards, Wesel, Germany), were grown in an RPMI medium with 10% fetal calf serum and 1% penicillin–streptomycin solution. All cells were cultivated in 5% CO₂ at 37 °C. Approximately 2.0 × 10⁶ cells were lysed in 100 µL lysis buffer (Tris–HCl 50 mM, EDTA 10 mM, NP-40 1%, SDS 1%) and physically fragmented by sonication to an average fragment length of 200–500 bp. The DNA was treated with 20 µg Proteinase K and purified using the NucleoSpin Gen and PCR Clean-up kit (Macherey–Nagel, Dueren, Germany). The purified DNA was kept at − 20 °C until applied to NGS.