Roche’s NGS Automation Support Team, in collaboration with non-Roche liquid handling vendors, creates menus of automated methods for KAPA library prep reagents. Let’s talk about the next steps to get you up and running with automating your NGS library prep protocols. Visit support for non-Roche liquid handlers to learn more.
KAPA EvoPrep Kits
For Research Use Only. Not for use in diagnostic procedures.
Streamline your workflow, boost your performance from mechanically fragmented DNA and cfDNA
Achieve enhanced NGS sample prep performance and confidence, even with difficult cfDNA and FFPET DNA inputs, with KAPA EvoPrep Boost Kit. This streamlined, automation-friendly workflow for NGS library prep of mechanically or naturally fragmented DNA includes an optimized amplification mix to ensure high-quality results for WGS, WES, and targeted sequencing applications.
- Increase efficiency and convenience with all-in-one enzyme and buffer ReadyMixes—available in either automation-friendly reagent plates or in tubes
- Recover more information with enhanced library conversion by the engineered KAPA EvoT4 DNA Ligase
- Maximize sequencing efficiency by ensuring high uniformity and fewer dropouts in difficult regions with an optimized low-bias PCR mix (KAPA EvoAmp ReadyMix)
- Cover regions of interest with depth and accuracy in target enrichment workflows
Increase efficiency and convenience
- Analyze a greater variety of samples with a wide range of DNA inputs (0.1 ng - 500 ng)
- Reduce hands-on-time and the number of reagent tubes with ReadyMix formulations, available in tubes or automation-friendly plates
- Shorten workflow time without compromising results with a 5-minute ligation time
Figure 1: Streamline DNA library prep from 0.1-500 ng of input DNA with KAPA Evo Prep Boost Kits’ ready-to-use master mixes.
Avoid reagent preparation with master mixes available in tubes or pre-plated.
Maximize sequencing efficiency
- Increase data confidence with reduced artifact rate
- Minimize coverage bias for highly uniform results and more cost-effective sequencing
Figure 2: KAPA EvoPrep Boost Kits deliver low-bias amplification, exceptional accuracy, and minimal artifacts for superior sequencing quality.
Whole genome sequencing libraries were prepared using 0.1 ng of DNA (B. pertussis) with the KAPA EvoPrep Boost Kit, Supplier N, Supplier Q, Supplier T and Supplier W, following each supplier’s instructions for use. (A) Lower amplification bias with the KAPA EvoPrep Boost Kits, resulting in more uniform coverage across the B. pertussis genome. Best balance of highest sequencing economy and result confidence observed with the KAPA EvoPrep Boost Kits by (B) Lowest error rate and (C) ultra- low artifact rate compared to other suppliers.
Cover regions of interest with depth and accuracy in target enrichment workflows
- Achieve high specificity and broadest target coverage at high sequencing depths
- Read deeper by recovering more unique molecules
Figure 3: Improved sequencing performance in target capture workflow utilizing cfDNA.
10 ng of cfDNA was used to prepare triplicate libraries with the KAPA EvoPrep Boost Kit or Supplier I, Supplier N, Supplier Q, Supplier T and Supplier W, following each supplier’s instructions for use. Libraries were enriched with the KAPA HyperCap Oncology Panel (214 Kb), following the KAPA HyperCap cfDNA Evolved workflow instructions1. The KAPA EvoPrep Boost Kit had the highest percentage of target coverage at ≥ 1000X compared to other suppliers and the highest unique depth compared to other suppliers, thereby showcasing the optimal utilization of sequencing throughput by higher uniformity; as well as highest genome equivalent recovery rate compared to Supplier N, Supplier Q, Supplier T and Supplier W, resulting in higher data confidence2.
Thinking of automating library prep on your existing liquid handler?
Data on file. All graphic data is on file, unless otherwise noted.
References
1. KAPA EvoPrep Boost Kit data generated in 2025, separate from other supplier data generated in 2024.
2. McNulty, et al. (2020). Impact of reducing DNA input on next-generation sequencing library complexity and variant detection. The journal of Molecular Diagnostics, Volume 22, Issue 5, May 2020, Pages 720-727.
Simplify high-performance library prep from mechanically fragmented DNA and cfDNA
Achieve high-performing NGS library preparation from mechanically fragmented DNA or cell-free DNA (cfDNA) in a streamlined, automation-friendly workflow with KAPA EvoPrep Kit. The KAPA EvoPrep workflow reduces pipetting and reagent preparation steps and is validated with challenging sample inputs such as cfDNA and FFPET DNA.
KAPA EvoPrep Kit is the next evolution of Roche DNA Library Preparation portfolio, delivering unparalleled convenience for whole-genome sequencing (WGS), whole-exome sequencing (WES) and targeted sequencing applications–including cfDNA applications.
- Increase efficiency & convenience with automation-friendly ReadyMix reagents and new KAPA EvoT4 ligase, in plated format or in tubes
- Achieve higher yields & greater library conversion efficiency across a range of sample types and DNA inputs as low as 100 pg
- Reduce sequencing artifacts & coverage bias
- Improve performance with challenging sample types such as cfDNA
- Cover regions of interest with depth & accuracy in target enrichment workflows
Increase efficiency & convenience
- Analyze a greater variety of samples with a wide range of DNA inputs (0.1 ng-500 ng)
- Reduce hands-on-time & the number of reagent tubes with ReadyMix formulations, available in tubes or automation-friendly plates.
- Shorten workflow time without compromising results with a 5-minute ligation time
Figure 1: KAPA EvoPrep ReadyMix reagents are ready-to-use; no master mix preparation is required. NOTES: KAPA full-length UDI adapters are only available in 96- well plates. KAPA Universal Adapters, which are used in combination with KAPA UDI Primer Mixes and require amplification, are available in plates or tubes. KAPA HyperPure Beads, UDI Adapter Kits & KAPA Library Amplification Primer Mix (10X) are sold separately.
Achieve higher yields & greater library conversion efficiency
- Reduce duplication rates while increasing sequencing coverage, with fewer amplification cycles needed for downstream processing
- Achieve successful library construction with clinically relevant samples & PCR-free workflows from as little as 50 ng
Figure 2: KAPA EvoPrep chemistry enables high library conversion across a range of input DNA. 0.1 ng-500 ng of Covaris-sheared human genomic DNA was used to prepare libraries with KAPA Universal Adapters (with KAPA UDI Primer Mixes) at the recommended adapter: insert molar ratio following the KAPA EvoPrep and KAPA HyperPrep Kit Instructions for Use. Data bars represent the mean, and error bars represent standard deviation (n=4). *Non-validated input (outside of the input range) for KAPA HyperPrep Kit—optimized cycle number for KAPA EvoPrep Kit inputs used.
Reduce sequencing artifacts & coverage bias
- Improve your sequencing metrics with reduced artifacts, allowing higher confidence in your data
- Minimize coverage bias, increasing uniformity of coverage & potentially decreasing sequencing costs
Figure 3: KAPA EvoPrep Kit yields uniform coverage with reduction in sequencing artifacts. PCR-free whole genome libraries were prepared using 100 ng of Covaris-sheared human genomic DNA (NA12878) with the KAPA EvoPrep Kit, Supplier Q & Supplier W, following each supplier’s instructions for use. (A) The KAPA EvoPrep Kit had the lowest percentage of Chimeras present. (B) The percentage of SSARs present in data generated with the KAPA EvoPrep Kit was equivalent to Supplier Q, with Supplier W having the highest percentage of SSARs present. SSARs represent chimeric reads that appear to be derived from non-contiguous portions of the genome.³ (C) The KAPA EvoPrep Kit had uniform coverage and low AT-or GC-bias while Supplier Q had lumpy coverage and coverage hotspots i.e., over-representation of AT-rich region. This could require more sequencing to be performed to achieve the requisite coverage for these regions, which increases cost and turnaround times. (D) KAPA EvoPrep Kit had very low duplication count compared to Supplier Q & Supplier W, thereby having more unique library molecules present1. All libraries were sequenced on an Illumina NovaSeq 6000 (2x150bp) & subsampled to 150 M reads.
Improve performance with challenging sample types such as cfDNA
- Generate more diverse libraries from challenging sample types such as cfDNA
- Increase confidence in data due to reduction in sequencing artifacts
Figure 4: KAPA EvoPrep Kit yields a high number of unique library molecules, with fewer chimeric reads. Whole genome sequencing libraries were prepared using 1 ng of cfDNA with the KAPA EvoPrep Kit, Supplier N, Supplier Q, Supplier T & Supplier W, following each supplier’s instructions for use. KAPA EvoPrep Kit generated a higher number of unique library molecules compared to Supplier N, Supplier Q, Supplier T & Supplier W, resulting in higher library diversity & data confidence1. The KAPA EvoPrep Kit generated a low percentage of Chimeras present, resulting in higher data confidence2, with Supplier N, Supplier Q & Supplier W generating a higher percentage of Chimeras present. All libraries were sequenced on an Illumina NovaSeq 6000 (2x150bp) & subsampled to 400 M reads.
Cover regions of interest with depth & accuracy in target enrichment workflows
- Achieve high specificity with the highest percent of target coverage at ≥1000x
- Combine high uniformity & high recovery of unique molecules
Figure 5: KAPA EvoPrep Kits yield improved sequencing performance in target capture workflows utilizing cfDNA. 10 ng of cfDNA was used to prepare triplicate libraries with the KAPA EvoPrep Kit, Supplier N, Supplier Q, Supplier T & Supplier W, following each supplier’s instructions for use. Libraries were enriched with the KAPA HyperCap Oncology Panel (214 Kb), following the KAPA HyperCap cfDNA Evolved workflow instructions. The KAPA EvoPrep Kit had the highest percentage of target coverage at ≥1000X compared to other suppliers; the highest percentage of coverage uniformity compared to other suppliers, thereby showcasing the optimal utilization of sequencing throughput by higher uniformity; as well as higher unique molecule recovery compared to Supplier N, Supplier Q, Supplier T & Supplier W, resulting in higher data confidence1. All libraries were sequenced on an Illumina NovaSeq 6000 (2x150bp) and subsampled to 150 M reads.
Thinking of automating library prep on your existing liquid handler?
Roche’s NGS Automation Support Team, in collaboration with non-Roche liquid handling vendors, creates menus of automated methods for KAPA library prep reagents. Let’s talk about the next steps to get you up and running with automating your NGS library prep protocols. Visit support for non-Roche liquid handlers to learn more.
- McNulty et al. Impact of reducing DNA input on next-generation sequencing library complexity and variant detection. The journal of Molecular Diagnostics, Volume 22, Issue 5, May 2020, Pages 720-727.
- Chen et al. Characterization and mitigation of artifacts derived from NGS library preparation due to structure-specific sequences in the human genome. BMC Genomics (2024) 25:227.
- Haile, et al. (2019). Sources of erroneous sequences and artifact chimeric reads in next generation sequencing of genomic DNA from formalin-fixed paraffin-embedded samples. Nucleic Acids Research, 2019, 7,2. doi: 10.1093/nar/gky1142.
For Research Use Only. Not for use in diagnostic procedures.
HYPERCAP, KAPA HYPERPETE, KAPA, KAPA EVOT4, KAPA EVOPREP & KAPA HYPERPREP are trademarks of Roche.
All other product names and trademarks are the property of their respective owners.
| Cap or Plate Color | Component Name | Included in KAPA EvoPrep Kits (10154039001) (10153806001) |
Included in KAPA EvoPrep Kits (VK*) (10212233702) (10212250702) |
Included in KAPA EvoPrep Kits (PCR-free) (10153849001) (10153857001) |
|---|---|---|---|---|
| Blue | End Repair and A-Tailing ReadyMix | Yes | Yes | Yes |
| Yellow | Ligation ReadyMix | Yes | Yes | Yes |
| Green | KAPA HiFi HotStart ReadyMix (2X) | Yes | Yes | No |
| Green | KAPA Library Amplification Primer Mix (10X) | No | Yes | No |
For Research Use Only. Not for use in diagnostic procedures.
HYPERCAP, KAPA HYPERPETE, KAPA, KAPA EVOT4, KAPA EVOPREP & KAPA HYPERPREP are trademarks of Roche.
All other product names and trademarks are the property of their respective owners.
Request a KAPA EvoPrep Kits evaluation
*KAPA EvoPrep evaluation offer is valid for U.S. based customers only.