All KAPA Adapter Kits contain KAPA Adapter Dilution Buffer and three additional sealing films to support multiple use. KAPA Adapter Dilution Buffer (08278539001) is also available separately, if required.
For Research Use Only. Not for use in diagnostic procedures.
The KAPA EvoPlus Kits offer improved fragmentation performance, insensitivity to fragmentation inhibitors and reduced sequencing artefacts through our streamlined and fully automatable workflow. Bringing a new and upgraded enzymatic fragmentation and library prep solution to the market will enable researchers to achieve higher confidence with increased sequencing efficiency.
The KAPA EvoPlus Kits offer a complete library preparation solution when combined with KAPA Adapters and KAPA HyperPure Beads (sold separately). The kits are compatible with the Illumina sequencing platform and have been qualified with automation methods.
View comparative data showing negligible sequencing artefacts for both KAPA EvoPlus Kits and KAPA HyperPrep Kits (mechanical fragmentation) compared to other suppliers.
Figure 1. Improved sequencing performance - reduction of Strand-split artefact reads (SSARs). Human whole genome libraries were prepared using 500 ng inputs (size selection performed) with the KAPA EvoPlus Kit, Vendor N and Vendor Q. The percentage SSARs present for KAPA EvoPlus libraries are equivalent to KAPA HyperPrep libraries - near zero (0.002%). The competitor kits have a higher percentage of SSARs present. SSARs represent chimeric reads that appear to be derived from non-contiguous portions of the genome.*
* Source: 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, 47,2. doi: 10.1093/nar/gky1142.
Figure 2. Improved sequencing performance - reduction of chimeras. Human whole genome libraries were prepared using 500 ng inputs (size selection performed) with the KAPA EvoPlus Kit, Vendor N and Vendor Q. The percentage chimeras present for KAPA EvoPlus libraries is similar to the KAPA HyperPrep libraries. The competitor kits have a higher percentage of chimeras present.
Figure 3. Improved sequencing performance - high positive predictive values (PPVs). Human whole genome libraries were prepared using 500 ng inputs (size selection performed) with the KAPA EvoPlus Kit, Vendor N and Vendor Q. The PPV for the KAPA EvoPlus libraries are similar to the KAPA HyperPrep libraries (considered industry gold-standard). The competitor kits have a lower PPVs.
Figure 4. The KAPA EvoPlus Kit offers a simplified and streamlined workflow to remove the complexities and risk for human error by providing a trusted enzymatic fragmentation solution.
See performance data showing tunability and consistency in DNA library insert sizes with samples from different input amounts and DNA dilution buffer.
Figure 5. The KAPA EvoPlus chemistry enables reproducible enzymatic fragmentation across DNA diluted in different buffer types. Human genomic DNA (500 ng) diluted in 10 mM Tris-HCl and 10 mM Tris-HCl, 1 mM EDTA was fragmented at 37°C to achieve mode library insert sizes of approximately 300 bp. The KAPA EvoPlus workflow was completed without any size selection using full-length adapters (KAPA UDI Adapters) and 2 cycles of amplification. Final libraries were analyzed using a LabChip GX Touch HT instrument and HT DNA HiSens Reagent Kit (PerkinElmer).
Figure 6. The KAPA EvoPlus chemistry enables reproducible enzymatic fragmentation across different input amounts. Human genomic DNA (10 ng, 100 ng and 500 ng) was fragmented at 37°C to achieve mode library insert size of approximately 300 bp. The KAPA EvoPlus workflow was completed without any size selection using full-length adapters (KAPA UDI Adapters) and 2 cycles of amplification. Final libraries were analyzed using a LabChip GX Touch HT instrument and HT DNA HiSens Reagent Kit (PerkinElmer).
Figure 7. The KAPA EvoPlus chemistry enables 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 workflow was completed without any size selection using full-length adapters (KAPA UDI Adapters) and 2 cycles of amplification. Final libraries were analyzed using a 2100 Bioanalyzer instrument and DNA High Sensitivity DNA Kit (Agilent).
The GC distribution for KAPA EvoPlus libraries are equivalent to KAPA HyperPrep libraries which use mechanically sheared DNA (considered industry gold-standard) as the input.
Figure 8. Enzymatic fragmentation in the KAPA EvoPlus workflow had no impact on overall coverage depth and uniformity, or GC bias - normalized coverage across the GC spectrum was virtually for KAPA EvoPlus and KAPA HyperPrep workflows. The Vendor Q workflow has challenges with both AT- and GC-rich genomes, whereas Vendor N workflow performs worse with GC-rich sequence— presumably as the result of more biased fragmentation and library amplification. This results in lumpy coverage and coverage hotspots, i.e., over-representation of "easy" (more GC-balanced) regions and under-representation of “difficult” (AT- and GC rich) regions. With these methods, more sequencing has to be performed to achieve the requisite coverage for these regions, which increases cost and turnaround times.
*Data on file.
All graphic data is on file, unless otherwise noted.
| Cap or plate colour | Component Name | Included in KAPA EvoPlus Kits (09420037001, 09420053001, 09420339001, 09420428001) |
Included in KAPA EvoPlus Kit (PCR-free) (09420045001, 09420304001, 09420371001, 09420436001) |
|---|---|---|---|
| Red | KAPA FragTail ReadyMix | Yes | Yes |
| Yellow | KAPA Ligation ReadyMix | Yes | Yes |
| Green | KAPA HiFi HS ReadyMix (2X) | Yes | No |
| Kit Code | Roche Cat. No | Description | Kit Size |
|---|---|---|---|
| N/A | 09420037001 | KAPA EvoPlus Kit | 24 rxn |
| N/A | 09420053001 | KAPA EvoPlus Kit | 96 rxn |
| N/A | 09420339001 | KAPA EvoPlus Kit | 384 rxn |
| N/A | 09420428001 | KAPA EvoPlus Kit, 96 well plate | 96 rxn |
| N/A | 09420045001 | KAPA EvoPlus Kit (PCR-free) | 24 rxn |
| N/A | 09420304001 | KAPA EvoPlus Kit (PCR-free) | 96 rxn |
| N/A | 09420371001 | KAPA EvoPlus Kit (PCR-free) | 384 rxn |
| N/A | 09420436001 | KAPA EvoPlus Kit (PCR-free), 96 well plate | 96 rxn |
All KAPA Adapter Kits contain KAPA Adapter Dilution Buffer and three additional sealing films to support multiple use. KAPA Adapter Dilution Buffer (08278539001) is also available separately, if required.
| Kit Code | Roche Cat. No | Description | Kit Size |
|---|---|---|---|
| KK8007 | 08963835001 | KAPA HyperPure Beads (5 mL) | 5 mL |
| KK8008 | 08963843001 | KAPA HyperPure Beads (30 mL) | 30 mL |
| KK8009 | 08963851001 | KAPA HyperPure Beads (60 mL) | 60 mL |
| KK8011 | 08963878001 | KAPA HyperPure Beads (4 x 60 mL) | 4 x 60 mL |
| KK8010 | 08963860001 | KAPA HyperPure Beads (450 mL) | 450 mL |
| KK8727 | 08861919702 | KAPA Unique Dual-Indexed Adapters Kit (15μM) | 96 adapters x 20 μL each |
| KK8721 | 08278539001 | KAPA Adapter Dilution Buffer (25 mL) | 25mL |
| N/A | 09063781001 | KAPA Universal Adapter, 15μM 960 μL | 96 samples |
| N/A | 09063790001 | KAPA Universal Adapter, 15μM 4x960 μL | 384 samples |
| N/A | 9329862001 | KAPA Universal UMI Adapter, 960 μL | 96 samples |
| N/A | 09329889001 | KAPA Universal UMI Adapter, 4x960 μL | 384 samples |
| N/A | 09134336001 | KAPA UDI Primer Mixes, 1-96, 96 rxn | 96 samples |
| N/A | 09329838001 | KAPA UDI Primer Mixes, 97-192, 96 rxn | 96 samples |
| N/A | 09329846001 | KAPA UDI Primer Mixes, 193-288, 96 rxn | 96 samples |
| N/A | 09329854001 | KAPA UDI Primer Mixes, 289-384, 96 rxn | 96 samples |