Scheme for laboratory process for the MRD-based SNP discovery. Individual PCR reactions from a homozygous source are pooled and cloned en masse to vector DNA to construct standards. Individual PCR reactions from the population of interest sample are pooled, hybridized to the standards' DNA, and transformed into the mutation sorter strain that enriches for the variant fragments. The resultant colonies are pooled, and the pool plasmid DNA is prepared followed by individual PCR and sequencing reactions for each amplicon. Sequencing traces from the enriched pool are compared with those of the homozygous source in order to detect SNP sites.
