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. 2021:2265:277-286.
doi: 10.1007/978-1-0716-1205-7_21.

Single-Cell Analysis of BRAFV600E and NRASQ61R Mutation Status in Melanoma Cell Lines as Method Generation for Circulating Melanoma Cells

Joseph W Po  1   2   3 , Yafeng Ma  4   5   6 , Alison W S Luk  4   7 , David Lynch  4   5   8 , Bavanthi Balakrishnar  9 , Daniel Brungs  4   5   10 , Farhad Azimi  9 , Adam Cooper  5   8   9 , Erin Saricilar  8   6   9   11 , Vinay Murthy  4   5   8   9 , Paul de Souza  4   8   6   9   12 , Therese M Becker  4   5   8   6
Affiliations

Single-Cell Analysis of BRAFV600E and NRASQ61R Mutation Status in Melanoma Cell Lines as Method Generation for Circulating Melanoma Cells

Joseph W Po et al. Methods Mol Biol. 2021.

Abstract

Molecular testing of tumor biopsies allows for the identification of the key mutations driving a patient's cancer. However, this is limited to singular nodes and may not accurately reflect cancer heterogeneity. Circulating tumor cell (CTC) analyses offer a noninvasive method of interrogating the genomic profile of patient-derived tumor material. To date, molecular analysis of CTCs has relied on the characterization of bulk or pooled CTC lysates, limiting the detection of minor tumorigenic CTC subclones. Here, we show a workflow enabling BRAFV600E/NRASQ61R mutation detection from single cultured melanoma cells by combining micromanipulation and genomic material amplification methods. This workflow can be directly integrated into circulating tumor cell analysis applications.

Keywords: Cell lines; Circulating tumor cells (CTC); Liquid biopsy; Melanoma; Whole-genome amplification (WGA).

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