DNA library preparation for next-generation sequencing (NGS) involves fragmentation of target DNA molecules into varying sizes, end repair or A-tailing and ligation of platform-specific adapters to the library. The libraries are then amplified and cleaned up subsequently. Depending on the nature of the DNA sample (e.g., fresh sample versus formalin-fixed paraffin embedded (FFPE) tissue), the workflow may require optimization.
Fragmentation of DNA can be achieved through mechanical or enzymatic shearing. While mechanical shearing is considered the gold standard, enzyme-based fragmentation affords speed, less hands-on time and convenience to enable automation of the workflow. Since the adapter size is constant, the library size is adjusted based on the desired insert size and the type of application. Amplification of library fragments is typically required for many applications, including target enrichment. Regions within the genome, for example those that may be GC-rich, are difficult to amplify and may introduce sequencing bias.
The KAPA EvoPrep Kits offer a streamlined and automation friendly library preparation solution, validated with challenging sample inputs such as cell-free DNA, FFPET DNA or mechanically sheared DNA. The KAPA EvoPlus V2 Kits offer improved fragmentation performance, insensitivity to inhibitors, better conversion efficiency and reduced sequencing artefacts, enabling researchers to achieve higher confidence with increased sequencing efficiency. KAPA EvoPrep and KAPA EvoPlus V2 Kits both feature ReadyMix reagents, reducing the number of tubes and workflow steps. Both kits are available in tube and plated formats for enhanced automatability. Designed to provide increased library conversion efficiency with the KAPA EvoT4 DNA Ligase in the Ligation ReadyMix, are designed to enable higher sensitivity and more confident variant detection.*
For targeted sequencing applications, our KAPA EvoPrep and KAPA EvoPlus V2 Kits can be combined with either the KAPA HyperCap or KAPA HyperPETE workflows to provide a complete and integrated sample prep solution.