KAPA Library Quantification Kits

Figure 4. High consistency across different library types and across hundreds of production lots. Nine human WGS libraries (41% GC) and two microbial WGS libraries (Rhodococcus sp., 70% GC and Staphylococcus sp., 35% GC) were quantified with distinct lots (Lots 1 and 2), and distinct sets of reagents from the same lot (Set 1 and 2) of KAPA Library Quantification Kit for the Illumina platform.

Figure 7. Accurate and sensitive quantification enables a uniform distribution of reads in muliplexed sequencing pools. Twenty-four indexed libraries, prepared from high-quality human genomic DNA (darker bars) or FFPE DNA (lighter bars), were quantified by qPCR using the KAPA Library Quantification Kit for Illumina platforms and combined to create three sequencing pools of equimolar concentration. Each pool was sequenced in a different lane of the same flow cell on a HiSeq 2500 instrument. (Figure 7) The twenty-four individual libraries represented a ~44-fold range of pre-pooling concentrations (5.2 nM to 229.8 nM), whereas the sequencing read distribution (Figure 8) only varied between 9.6% and 13.9%. The coefficient of variation (CV) for pre-pooling library concentration was 94.9%, 49.4%, and 127.9% for the libraries pooled for sequencing in lanes 1, 2, and 3, respectively. The KAPA Library Quantification Kit enabled accurate equimolar pooling, reducing the CV to 2.5%, 4.8%, and 11.2% after normalization.