Application: Bacterial Whole-Genome Sequencing Data on file. *Automation and Connectivity solutions are in development. KAPA and LightCycler are For Research Use Only. Not for use in diagnostic procedures. Solutions KAPA HyperPlus Kits with KAPA HiFi HotStart ReadyMix offer rapid, flexible and low-bias library construction, from extracted DNA or crude bacterial lysates. KAPA Dual-Indexed Adapters are QC-tested for NGS performance and barcode cross-contamination. KAPA Pure Beads offer efficient, tunable reaction cleanups. KAPA Library Quantification Kits enable accurate, qPCR-based library quantification. Compatible with Roche LightCycler ® 96 and LightCycler ® 480 real-time PCR systems. Application Challenges • Bacteria have very diverse cellular and genomic architecture. Robust methods are needed to process a wide range of species and strains in standardized sequencing pipelines. • Liquid culture of bacterial isolates and DNA extraction for WGS library preparation are time consuming and may add cost to the workflow. • Many important pathogens have AT- or GC-rich genomes. Biases introduced during sample preparation and sequencing makes it difficult to obtain reliable coverage of certain genomic regions or genes of interest. • De novo assembly from short-read sequencing technologies is inherently difficult, and more so when contigs are short and large stretches of genomic content are missing from sequence data. • Bacterial WGS is often peformed in high-throughput settings that require flexibility and quick turnaround times. Benefits • Rapidly prepare high-quality WGS libraries from extracted DNA, or directly from crude bacterial lysates to significantly reduce the turnaround time from bacterial isolate to sequence. • Eliminate the need for mechanical fragmentation with the KAPA HyperPlus Kit with integrated enzymatic fragmentation. Increase thoughput with a flexible, automation-ready protocol, that can be tailored to produce insert sizes that are optimal for your sequencing read length. • Introduce minimal bias during fragmentation and library amplification with KAPA Frag and KAPA HiFi enzymes. Higher uniformity typically translates to better coverage of difficult genomic regions, and longer contigs for de novo assembly. • Enjoy greater peace of mind with integrated support and service for a complete bacterial WGS workflow solution. Sample Sequencing Ready Library Automation & Connectivity S a m p l e C o ll e c t i o n S a m p l e E n r i c h m e n t N u c l e i c A c i d E x t r a c t i o n S a m p l e Q u a n t i fi c a t i o n / Q C L i b r a r y P r e p a r a t i o n T a r g e t E n r i c h m e n t L i b r a r y Q u a n t i fi c a t i o n * GC-bias plot for Clostridium difficile (29% GC) whole-genome libraries, prepared with the KAPA HyperPlus Kit (green), or kits from other suppliers (red and orange). With the Roche Sample Prep Solution, very even coverage was achieved across all regions of the AT-rich genome. Libraries prepared with other sample prep methods are missing more genomic content regions, and must be subjected to additional sequencing to produce the same average coverage. Normalized Coverage GC% of 100 Window Bins 40 45 50 10 35 30 25 20 15 0.0 0.5 1.0 1.5 2.0 2.5 3.0 Number of Windows (x 10 3 ) 300 250 200 150 100 50 0 Roche Sample Prep Workflow