Overview

As next-generation sequencing (NGS) sample preparation entails a multitude of steps and each step is critical for the overall sequencing success and efficiency, all steps need to be optimized in terms of reagents and protocol. Considering the precious nature of NGS samples, an integrated workflow with optimized reagents is necessary for converting the samples into a sequencing-ready library. NGS sample types can differ in terms of their:

  • Ease of sequencing - uniform or with secondary structures or GC-rich areas
  • Quality - freshly extracted or preserved (formalin-fixed paraffin-embedded (FFPE))
  • Volume available - reasonable amounts to very small sample volumes


Taking into account all these variations, it is important to use library preparation reagents that can work together with all these sample types. High-quality library preparation reagents help in obtaining uniform and improved coverage as well as lower duplication rates. Depending on the end goal (for example, whole exome sequencing), a target enrichment step may also be required for enriching specific regions of interest. Use of high-efficiency target enrichment reagents helps in obtaining uniform and high depth of coverage.

The latest version KAPA HyperCap Workflow combines validated products for both KAPA Library Preparation and KAPA HyperCap Target Enrichment Probes and all accessories and reagents needed for sample preparation into a single workflow. The new KAPA HyperCap Target Enrichment Probes provide high capture efficiency, sequencing uniformity and low duplication and together with the updated KAPA HyperPlus Kits, the workflow delivers better target coverage and higher variant detection confidence. In addition, the workflow includes KAPA Universal Adapter, KAPA Universal UMI Adapter and KAPA UDI primer mixes (up to 384). The KAPA HyperCap Workflow utilizes the renowned probe design and selection algorithm and provides the convenience of simplified ordering and support from a single vendor, and is automation friendly. Sequencing data generated with the KAPA HyperCap Workflow can be analyzed with Roche's cloud-based secondary analysis solution - NAVIFY® Mutation Caller - for accurate detection of germline SNV and indel variants.

Features and Benefits of the KAPA HyperCap Workflow

  • Higher library conversion efficiency and lower index misassignments with the innovative adapters and UDI primer mixes
  • Enhanced sequencing performance and a faster workflow with a new end-repair, A-tailing chemistry and a single-sided post-ligation size selection
  • Increased capture uniformity and sequencing efficiency with higher fidelity probes manufactured using KAPA HiFi polymerase
  • Confirmed presence and expected concentration of probes through NGS-based QC
  • Convenient all-inclusive workflow with validated products and simplified ordering and support from a single vendor
  • Confident germline SNV/Indel calling with NAVIFY® Mutation Caller

Integrated and streamlined workflow

  • Higher efficiency in library preparation process with lower input amounts
  • Higher coverage with low duplicate rates
  • Automation-friendly
  • Ordering and support of all products of the workflow from a single vendor
  • Bead-based hybridization setup, single temperature hybridization washes, UEO's included, no SpeedVac required

Universal Enhancing Oligos (UEOs) allow for easy experiment planning

  • Reduced time and complexity with the elimination of adapter-matched blocking oligo use
  • Flexibility to block 6- and 8-base barcode adapters and use with single- or dual-indexing strategies

Efficient library construction with KAPA HyperPlex Adapters

  • Minimal levels of adapter dimer formation
  • Nominal levels of barcode cross contamination as confirmed by sequencing-based QC testing
  • Higher multiplexibility compared to existing options
  • Higher library conversion rate
  • Accurate molecule counting and PCR/Sequencing error correction with the KAPA Universal UMI Adapter

Easily automatable workflow with broad platform compatibility 

  • Reduced instrumentation requirement enabling easy automation
  • Multiplexing option, optimizing robot utilization
  • Concordant performance as the manual method