Well Plate Lineage Sequencing for Ultra-High-Quality SNV and Indel Detection with Single Cell Cycle Resolution In Vitro

Lineage sequencing uniquely measures somatic single nucleotide variants in dividing cells to achieve high specificity/sensitivity as well as the ability to temporally resolve variants and to relate sequenced genotypes to optically observed cellular phenotypes. Despite the high-quality data and unique capabilities offered by this method, it had previously been underused due to a need for complex, microfluidic-based cell collection. We demonstrate novel protocols for performing lineage sequencing that enable easy adoption of the method without the need for highly specialized equipment or expertise. In addition, we expand the repertoire of mutations measurable with the technique to include variants that arise specifically in response to a genotoxic treatment. These proof of concept demonstrations were performed in the HeLa cell line.]]>