Figure 1. KAPA Universal Adapter with all 384 KAPA UDI Primer Mixes perform consistently in the KAPA HyperCap Workflow v3.
High reproducibility was demonstrated across all UDI pairs following the sequencing of duplicate libraries containing all 384 UDI pairs in a single run. Filtered reads delivered high specificity (% reads on-target), deep target coverage (% bases covered by > 50X), and high uniformity (% bases within 0.5X – 2X of median coverage). Total duplicate rate was 3.2 % + 0.2 % and fold-80 base penalty was 1.32 + 0.01. Experimental design: Duplicate libraries were prepared with the KAPA HyperCap Workflow v3 using the KAPA HyperCap Heredity Panel (10 Mb capture target) and the KAPA HyperPlus Kit; input = 100 ng of human genomic DNA (NA12878; Coriell Institute). Libraries were multiplexed prior to capture, a total of 12 x 32-plex captures (384 enriched libraries total). All enriched libraries were then pooled and sequenced on a NovaSeq™ 6000 System lane at 2 x 100 bp, resulting in a mean of ~56.8 Million reads per sample after quality filtering. After down-sampling at 20 Million reads per sample, analysis followed the technical note “How To Evaluate KAPA Target Enrichment Data” (March 2020)1. Total duplicate rate was 3.2 % + 0.2 % and fold-80 base penalty was 1.32 + 0.01.
1. Meyer, J, et al. Roche Application Note SEQ100183. 2018.