Report on the optimum protocol for ctDNA isolation and enrichment

): Outcomes from the previous tasks will be combined and two alternative options will be explored. In the first and simplest scenario, captured ctDNA on beads will be directly mixed with the 2 pairs of oligos and the ligase chain reaction will be performed in situ, while the second scenario involves release of captured ctDNAs from the beads and subsequent mixing with the LCR cocktail. In both cases, contaminants are removed, serum is exchanged to buffer and ctDNA targets are specifically captured and enriched at least to the aM range. The protocol will be developed using synthetic targets: a) in buffer, b) spiked-in serum.