APPLICATION NOTE MagMAX Cell-Free DNA Isolation Kit Extraction and analysis of circulating cell-free DNA from plasma samples for lung cancer research

Lung cancer is a leading cause of death worldwide, with the majority of cases attributed to non-small cell lung carcinoma [2]. Mutation of the epidermal growth factor receptor gene (EGFR) is frequently observed in non-small cell lung carcinoma. For lung cancers positive in this mutation, first-generation EGFR tyrosine kinase inhibitors (EGFR-TKIs) are applied, which often have a remarkable effect, but eventually develop tolerance [3]. The T790M mutation of EGFR is recognized in approximately half of drug-resistant tumors, and third-generation EGFR-TKIs are effective against lung cancers with this mutation. Before treatment with targeted therapies, it is essential to confirm the T790M mutation using companion diagnostics. As previously described, it is often difficult to perform a second biopsy of the tumor tissue, so genetic testing using cfDNA from peripheral blood is of interest to many researchers studying practical methods to replace a tissue biopsy. Abstract We report a method for the extraction of circulating cell-free DNA (cfDNA) from plasma samples using the Applied BiosystemsTM MagMAXTM Cell-Free DNA Isolation Kit followed by digital PCR and next-generation sequencing analysis. Extraction and analysis of cfDNA was successfully performed from the plasma of 3 donors in good health and 11 donors with lung cancer. The results show that the MagMAX Cell-Free DNA Isolation Kit offers efficient recovery of high-quality cfDNA for downstream research applications and enables time savings compared to traditional methods.