Figure 1.

SMRT sequencing pipeline. Each amplicon was barcoded with unique forward and reverse primer pairs in a 96-well format. Complementary amplicon strands are dark and light blue bars; black and red bars correspond to forward and reverse barcodes, respectively, conjugated to target-specific primers (arrows). Following amplicon pooling, hairpin adapters (orange) were attached during SMRT library construction, converting linear into circular molecules. A single DNA polymerase (purple) reads each circular template in real time during the sequencing reaction. Circular consensus sequence (CCS) reads obtained from a SMRT cell were sorted by barcodes and aligned to a reference sequence.