Fig. 2From: Application of nonsense-mediated primer exclusion (NOPE) for preparation of unique molecular barcoded librariesExemplary experiment. DNA library construction and neutralization of UMI after PCR-based UMI introduction. Genomic DNA samples are amplified using 10Â cycles of linear PCR with EGFR exon 20 specific primer modified to include UMI and partial Illumina sequencing adapter. Remaining UMI-primer is neutralized using 10Â cycles of annealing/elongation in the presence of NOPE oligo. Next, 3 rounds of PCR are performed for amplification of EGFR gene fragments and introduction of complete Illumina adapters and indexes sequencesBack to article page