A Droplet PCR-Based Next Generation Sequencing Assay to Track Plasma DNA Mutation Dynamics in Estrogen Receptor Positive Metastatic Breast Cancer

Here we describe a protocol that uses droplet polymerase chain reaction for target enrichment followed by next generation sequencing of blood plasma circulating tumor DNA. This technique was used to characterize mutations in the genes ESR1 (all coding regions), TP53 (all coding regions), PIK3CA (hotspots), PIK3R1 (hotspots) and POLE (exonuclease domain).