Detection of MET and EML4-ALK mutations

RNA was retrotranscribed with the M-MLV retrotranscriptase (ThermoFisher Scientific, Waltham, MA, USA) and random primers. HotStart Taq polymerase (Qiagen) was used for METΔex14 and EML4-ALK amplification using a 20 µL reaction (45 cycles) and visualized in agarose gels, as described [9 (link),11 (link)]. Positive samples were confirmed by bidirectional Sanger sequencing of RT-PCR products, using the big-dye 3.1 sequencing kit (Applied Biosystems, Foster City, CA, USA).

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Aguado C., Giménez-Capitán A., Román R., Rodríguez S., Jordana-Ariza N., Aguilar A., Cabrera-Gálvez C., Rivas-Corredor C., Lianes P., Viteri S., Moya I, & Molina-Vila M.A. (2020). RNA-Based Multiplexing Assay for Routine Testing of Fusion and Splicing Variants in Cytological Samples of NSCLC Patients. Diagnostics, 11(1), 15.

Publication 2020

M-MLV retrotranscriptase HotStart Taq polymerase big-dye 3.1 sequencing kit

Agarose Gels Primers Rt pcr Taq polymerase

Corresponding Organization : Hospital Universitario Dexeus

Other organizations : Instituto Oncológico Dr. Rosell, Hospital de Mataró, Hospital General de Catalunya

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression of MET∆ex14 and EML4-ALK

control variables

None explicitly mentioned

controls

Positive controls: None mentioned
Negative controls: None mentioned

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