Abstract

Novel Methods for Enrichment of Mutations and Differentially Methylated Sequences from Circulating DNA

Mike Makrigiorgos, Professor, Harvard Medical School

Circulating DNA is poised to become a widely used tool for repeated assessment of cancer mutation and methylation status during the course of therapy. Removing the high excess wild type DNA fraction from circulating DNA allows enrichment of variant DNA and boosts the potential of all endpoint detection technologies, including sequencing. We present Nuclease-assisted Mutation Enrichment with Probe Overlap, NaME, a simple and powerful approach to remove wild type DNA from large gene pools simultaneously, in order to focus sequencing on clinically relevant DNA alterations. This single-step approach retains current sample preparation protocols almost unchanged and combines seamlessly with downstream technologies such as HRM, COLD-PCR, ddPCR and next generation sequencing. Application in clinical samples and liquid biopsies will be presented.