Single-molecule mutation sequencing of metallothionein 2A suppressed cadmium treated ovarian cancer cells

CAOV3 ovarian cancer cells were passaged in the presence or absence of low dose cadmium chloride for 15 passages. Cells were knocked down with shRNA targeting MT2A or a scrambled control vector. At the completion of passaging, genomic DNA was isolated. Random genomic DNA was assessed for mutations using single-molecule mutation sequencing, a method utilizing rolling circle amplification of DNA duplexes to discover and quantify ultra-low frequency random mutations.