Duplex Proximity Sequencing (Pro-Seq): A method to improve DNA sequencing accuracy without the cost of molecular barcoding redundancy
In many cases, errors are corrected automatically on the sequencer as they represent a minority sequence compared to the dominant base within a cluster, and are ignored or not detected by the basecaller. To check for errors (mixed bases) that are of similar frequency to the correct base, the relative fluorescence (fQ) is calculated for each base in a read, in such a way that dips represent the presence of a mixed base. An adjustable threshold is used to identify dips and only the mixed base position is then masked. The rest of the read can be trusted to provide high-fidelity sequence information.