KAPA EvoPlus Kits

• Produce sequencing- or target capture-ready libraries in ~2  hours
• Reduce hands-on time and opportunity for error with fewer enzymatic steps, ready-to-use (ReadyMix) reagent formulations, and fewer pipetting steps
• Automate the entire protocol from sample to library, including tunable enzymatic fragmentation
• Choose from tube-based or plated reagents, whichever best suits your laboratory setup and throughput or project size
• Offer a complete library prep solution when combined with KAPA adapters and KAPA HyperPure Beads (sold separately)
• Suitable for whole genome sequencing of any sized genome, and validated with KAPA HyperCap and KAPA HyperPETE target enrichment workflows and probes to support whole exome sequencing and targeted sequencing

Figure 1: The KAPA EvoPlus workflows and reagents 

KAPA EvoPlus Kits provide highly streamlined, automation-friendy workflows to save time and improve efficiency. KAPA EvoPlus Boost Kits include an amplification module with the KAPA EvoAmp ReadyMix powered by HiFi. 

• Adjust library insert size simply by varying fragmentation time
• Reproducible insert sizes across a range of DNA input amounts and GC contents 
• Robust and reliable results with real-life samples, such as those  in buffers containing EDTA (up to 2 mM) and other inhibitors commonly found in extracted DNA.

Figure 2: Tunable enzymatic fragmentation

Human genomic DNA (100 ng) was fragmented at 37°C for different periods of time (5 – 30 min) to achieve mode library insert sizes ranging from approximately 250 – 1000 bp. The KAPA EvoPlus V2 workflow was completed using full-length KAPA UDI Adapters and 3 cycles of amplification to enable visualisation. Electropherograms were generated with LabChip GX Touch NGS 3K Assay.

Figure 3: Reproducible enzymatic fragmentation in different buffer types

High-quality human genomic DNA (100 ng), eluted in 10 mM Tris-HCl, pH 8.0, 10 mM Tris-HCl, pH 8.0 + 1 mM EDTA, or PCR-grade water was fragmented for 15 min and converted to sequencing-ready libraries with the KAPA EvoPlus V2 Kit. KAPA UDI Adapters were used at the recommended adapter: insert molar ratio, as outlined in the KAPA EvoPlus V2 Kit Instructions for Use. Libraries were amplified for 3 cycles to enable visualization. Electropherograms were generated with LabChip GX Touch NGS 3K Assay.

• Highly efficient conversion of input DNA to library molecules with the engineered KAPA Evo T4 Ligase increases library yields and success rates from low inputs and difficult samples, in both PCR-free WGS and targeted sequencing workflows¹
• The optimized KAPA EvoAmp ReadyMix powered by HiFi, delivers exceptional coverage uniformity and extremely low error rates. This ensures high-confidence results and less sequencing to adequately cover challenging genomic regions.
• Highly optimized, low-bias enzymatic fragmentation and library amplification minimize sequencing artifacts (e.g., chimeras, soft-clipped reads, strand-split artifact reads) and associated wasted bases/reads.

Figure 4: Low-bias amplification, exceptional accuracy, and minimal artifacts support superior sequencing quality and economy.

Whole genome sequencing libraries were prepared from 100 pg inputs of Clostridium difficile genomic DNA, using the KAPA EvoPlus Boost Kit with KAPA EvoAmp ReadyMix or comparable kits from five other suppliers. All libraries were prepared according to the supplier’s instructions for use.

(A) Lower amplification bias with the KAPA EvoPlus Boost Kit, resulted in highly uniform coverage across the AT-rich C. difficile genome. Furthermore, KAPA EvoPlus Boost Kits ensure high-confidence results and excellent sequencing economy through a combination of (B) the lowest error rate, and (C) ultra-low artifact rate, compared to kits from suppliers.

• Despite obvious workflow advantages, broad-based adoption of enzymatic fragmentation has been hampered by sequencing artifacts produced by first-generation enzymatic fragmentation chemistries. The advanced KAPA EvoPlus FragTail chemistry was specifically designed to minimize these artifacts and boost confidence in results.
• KAPA EvoPlus Kits perform similarly or better than kits that rely on mechanical fragmentation, particularly with challenging samples such as FFPE tissue DNA, across a broad range of library quality, sequencing, and target enrichment metrics.

Figure 5. Workflow efficiency meets industry-leading performance.

Low-quality FFPE tissue DNA was used to prepare triplicate libraries, using the KAPA EvoPlus Boost Kit with enzymatic fragmentation. The same DNA sample was mechanically sheared and used for library preparation with kits from five other suppliers. Each supplier’s instructions for use were followed. All libraries were enriched with the KAPA HyperCap Oncology Panel (214 Kb), as outlined in the KAPA HyperCap FFPET Evolved Workflow Instructions for Use.*

(A) The KAPA EvoPlus Boost Kit outperformed all other kits in terms of specificity (expressed as percentage of reads on target). Other metrics including superior coverage of target bases (B), the lowest percentage of reads associated with artifactual chimeric reads (C ), and significantly better duplex recovery (D) confirm that KAPA EvoPlus Kits produce high-confidence data² and enable improved sequencing economy.

*KAPA EvoPlus Boost Kit data generated in 2025, separate from other supplier data generated in 2024.

• Innovative formulations and kit configurations significantly reduce the number of individual reagent containers and pipetting steps compared to competitor products and older generation Roche kits. This reduces plastic waste and packaging materials.
• KAPA adapters and purification beads are sold separately to support operational flexibility and reduce wastage
• All reagents included in KAPA EvoPlus Boost Kits are REACH* compliant

*Registration, Evaluation, Authorisation and Restriction of Chemicals