KAPA HyperExome

Figure 1. KAPA HyperExome yields greater % reads-on-target, deeper median coverage, and broader target coverage compared to the Supplier X Exome.

For both the KAPA HyperExome and Supplier X exome workflows: DNA from 16 cell lines was processed in triplicate (48 total libraries per workflow); input DNA was enzymatically sheared; samples were pre-capture multiplexed in sets of 8 and hybridized for 16 hours; final post-capture libraries were amplified with 8 PCR cycles; and libraries were sequenced (2 x 100 bp) on an Illumina® NovaSeq™ sequencer. For the KAPA HyperExome samples, libraries were prepared from 100 ng of DNA with KAPA HyperPlus Kits using KAPA Universal Adapter and KAPA UDI Primer Mixes; hybridization and washes were carried out at 55ºC following 8 pre-capture PCR cycles. Supplier X samples were prepared according the manufacturer’s instructions from 50 ng of genomic DNA. For analysis, sequencing data was subsampled proportionally to exome panel size to achieve the same targeted average depth of coverage.

Figure 2. KAPA HyperExome achieves highly uniform coverage—even across regions with high or low %GC. 

Prior to capture with KAPA HyperExome probes, libraries were prepared from 100 ng of DNA with KAPA HyperPlus Kits using KAPA Universal Adapter and KAPA UDI Primer Mixes. Libraries (8) were multiplexed prior to capture, hybridized to probes for 16 hours, amplified post-capture, and sequenced on an Illumina NovaSeq sequencer (2 x 100 bp). Bars represent data from 16 different cell lines processed in triplicate. Each dataset in Panel B is derived from a different cell line.

Figure 3. KAPA HyperExome provides better coverage of important genomic databases compared to the Supplier X Exome. 

For both the KAPA HyperExome and Supplier X exome workflows: DNA from 16 cell lines was processed in triplicate (48 total libraries per workflow); input DNA was enzymatically sheared; samples were pre-capture multiplexed in sets of 8 and hybridized for 16 hours; final post-capture libraries were amplified with 8 PCR cycles; and libraries were sequenced (2 x 100 bp) on an Illumina® NovaSeq™ sequencer. For the KAPA HyperExome samples, libraries were prepared from 100 ng of DNA with KAPA HyperPlus Kits using KAPA Universal Adapter and KAPA UDI Primer Mixes; hybridization and washes were carried out at 55ºC following 8 pre-capture PCR cycles. Supplier X samples were prepared according the manufacturer’s instructions from 50 ng of genomic DNA. For analysis, sequencing data was subsampled proportionally to exome panel size to achieve the same targeted average depth of coverage.