Duplex Proximity Sequencing (Pro-Seq): A method to improve DNA sequencing accuracy without the cost of molecular barcoding redundancy
(A) Common molecular barcoding/UMI methods involve uniquely labeling each DNA molecule with a molecular identifier or barcode. Many copies of each barcoded molecule are sequenced, and reads from individual fragments are collected in software. True variants should be common to every read, while errors should only occur in a smaller fraction of the reads. 20 or more reads are often required to generate a software consensus for a single low-error read. (B) Pro-Seq physically links copies of the same starting fragment into a single complex. Each linked complex is then sequenced in a single cluster, producing a high-fidelity read without redundancy. Two to roughly 100 linkers have been tested with Pro-Seq, but four are shown for simplicity.