Patent Document ID: 10152569
Application ID: 14346954
Patent Status: 1

Claim One:
1. A method of developing a consensus sequence from a plurality of sequencing reads of a nucleic acid target, comprising: (i) ligating a target nucleic acid and an anchor segment of known sequence and thereby forming a nucleic acid target template, which is a circular molecule in which the target nucleic acid forms an adjacent segment adjacent the anchor segment, (ii) generating raw target sequencing reads by synthesis directed by a polymerase reading around the nucleic acid target template multiple times primed from a primer binding to the anchor segment, the raw target sequencing reads comprise alternating reads of the anchor segment and the target nucleic acid; and at least some of the raw target sequencing reads containing sequencing errors; (iii) performing the following computer-implemented steps: (a) receiving a population of raw target sequencing reads of the nucleic acid target template ; (b) evaluating the accuracy of sequencing of the anchor segment in different raw target sequencing reads by comparing raw target sequencing reads of the anchor segment with the known sequence of the anchor segment; (c) assigning a subset of the raw target sequencing reads into an accepted class based on reaching at least a threshold level of accuracy of the sequencing of the anchor segment; (d) polling nucleobases at a position equidistant to the anchor segment sequence in raw target sequencing reads in the accepted class to determine a consensus nucleobase, which consensus nucleobase is assigned as the first nucleobase of a nascent sequence of the adjacent segment; (e) assigning raw target sequencing reads having the consensus nucleobase determined in the prior polling step to remain in the accepted class and assigning raw target sequencing reads lacking the consensus nucleobase determined in the prior polling step to a rejected class; (f) optionally reassigning a raw target sequencing read from the rejected class to the accepted class by scoring similarity of the raw target sequencing read to the nascent sequence and reintroducing the raw target sequencing read if the sequence similarity reaches at least a threshold level of similarity; and (g) repeating steps (d), (e) and optionally (f), except that a repetition polls a position adjacent the position poled in the previous polling step for raw target sequencing reads having the consensus nucleobase polled in the previous step or in the case of a raw target sequencing read reassigned from the rejected class to the accepted class and not polled in the previous polling step or if polled and not having the consensus nucleobase in the previous polling step, the polling polls a position adjacent the position aligned with the last nucleobase of the nascent sequence to determine a consensus nucleobase, and the consensus nucleobases determined in successive repetitions are assigned as successive nucleobases in the nascent sequence of the adjacent segment; wherein step (f) is performed at least once.