For purchasing or general questions
For Research Use Only. Not for use in diagnostic procedures.
KAPA EvoPlus is a new enzymatic DNA library preparation solution designed to rival the gold-standard performance of mechanical-shearing methods while delivering unparalleled convenience for whole-genome (WGS), whole-exome (WES), and targeted sequencing applications performed on Illumina sequencing systems.
While enzymatic fragmentation is favored for the gains in efficiency, minimized sample loss, and ease of scalability that the method affords over mechanical-shearing, it has yet to achieve the same level of data quality. Enzymatic approaches often produce sequencing artifacts that obscure usable findings, thus forcing labs to trade performance for convenience. KAPA EvoPlus is an evolution in enzymatic library preparation that brings new levels of performance to enzymatic fragmentation, allowing labs to realize even greater convenience without sacrificing performance.
Figure 1: Comparison of the KAPA EvoPlus workflow to another enzymatic fragmentation kit supplier (“Supplier A”). Both workflows are carried out in a single tube up to the optional library amplification step. KAPA EvoPlus ReadyMix reagents are ready-to-use; no master mix preparation is required. With KAPA EvoPlus, only two reagents (ReadyMix #1 and ReadyMix #2) are added to input DNA vs. the Supplier A workflow which requires additional reagents and mixing steps, thus, increasing the quantity of consumables used and the number of pipetting steps. The core reaction (all non-optional steps) for EvoPlus can be completed in only four pipetting steps vs. Supplier A which requires nine.
Figure 2: KAPA EvoPlus yields reproducible fragment sizes with input DNA of varying quality. Experimental details: For low-and medium-quality input, DNA was extracted from clinical FFPET and fresh-frozen tissue using the KAPA NGS DNA Extraction Kit; no buffer exchange was performed prior to library preparation. For high-quality samples, Coriell DNA (NA12878) was used as input. For all samples: input into library prep was 100 ng DNA; fragmentation was carried out for 20 min; and KAPA Universal Adapters, KAPA UDI Primer Mixes, and KAPA HyperPure Beads were used.
Figure 3: KAPA EvoPlus chemistry yields consistent DNA fragmentation in a wide variety of buffers. For each condition tested, 100 ng of intact hgDNA was suspended in a different elution buffer (a total of 10 buffers) and subjected to the following experimental conditions: 25 min fragmentation at 37°C; adapter ligation with 15 µM KAPA UDI adapters; and eight PCR amplification cycles.
Figure 4: KAPA EvoPlus chemistry enables reproducible enzymatic fragmentation across a range of different input amounts. Human genomic DNA (10 ng, 100 ng, 500 ng) was fragmented at 37°C to achieve a 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).
Sequencing artifacts alter the true mutation signature of samples and reduce sequencing coverage and uniformity, leading to additional sequencing runs, inflated costs, and the investment of more effort to realize actionable results. KAPA EvoPlus outperforms leading enzymatic fragmentation kits, resulting in reduced artifacts—even delivering results comparable to gold-standard mechanical shearing. With KAPA EvoPlus, labs can act with confidence on their results.
Figure 5: The positive predictive values (PPV) obtained with KAPA EvoPlus are similar to mechanical-shearing.PPV refers to the proportion of positive test results that are truly positive. A higher PPV correlates with higher precision. Experimental conditions: human whole genome libraries were prepared using 500 ng hgDNA and either the KAPA HyperPrep Kit (mechanical fragmentation) or, for enzymatic fragmentation, KAPA EvoPlus, Supplier A, or Supplier B—each according to manufacturer’s instructions. Double-sided size selection was performed in all cases.
Figure 6: Detection of strand split artifact reads (SSARs) with KAPA EvoPlus were undetectable and comparable to mechanical shearing. The percentage of SSARs present in supplier kits represent chimeric reads that appear to be derived from non-contiguous portions of the genome. Human whole genome libraries were prepared using 500 ng hgDNA and either the KAPA HyperPrep Kit (mechanical fragmentation) or, for enzymatic fragmentation, KAPA EvoPlus, Supplier A, or Supplier B—each according to manufacturer’s instructions. Double-sided size selection was performed in all cases.
Figure 7: Coverage and uniformity achieved by KAPA EvoPlus was virtually identical to KAPA HyperPrep (mechanical fragmentation). Other technologies demonstrated decreased uniformity and coverage as the %AT- or GC-rich content increased. A normalized coverage score of 1.0 is reflective of uniform coverage across the range of %AT/GC content.
For purchasing or general questions
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| 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 |
For purchasing or general questions
For online ordering
| Roche Cat. No |
Description |
Kit Size |
How to buy |
|---|---|---|---|
| 9420037001 |
KAPA EvoPlus Kit, 24 rxn |
24 rxn |
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| 9420045001 |
KAPA EvoPlus Kit, 24 rxn (PCR-free) |
24 rxn |
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| 9420053001 |
KAPA EvoPlus Kit, 96 rxn |
96 rxn |
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| 9420304001 |
KAPA EvoPlus Kit, 96 rxn (PCR-free) |
96 rxn |
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| 9420428001 |
KAPA EvoPlus Kit, 96 rxn plate |
96-well Plate |
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| 9420436001 |
KAPA EvoPlus Kit, 96 rxn plate (PCR-free) |
96-well Plate |
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| 9420339001 |
KAPA EvoPlus Kit, 384 rxn |
384 rxn |
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| 9420371001 |
KAPA EvoPlus Kit, 384 rxn (PCR-free) |
384 rxn |
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| Roche Cat. No |
Description |
Kit Size |
How to buy |
|---|---|---|---|
| 07958927001 |
HotStart ReadyMix, 100 x 25 µL reactions | 1.25 mL |
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| 07958935001 |
HotStart ReadyMix, 500 x 25 µL reactions | 6.25 mL |
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| 09420398001 |
HiFi HotStart ReadyMix, 9.6ml |
9.6 mL |
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| Roche Cat. No |
Description |
Kit Size |
How to buy |
|---|---|---|---|
| 07958994001 |
KAPA Library Amp Primer Mix | 250 reactions |
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| 09420410001 |
KAPA Library Amp Primer Mix (384 rxn) |
384 rxn |
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| 09420479001 |
KAPA Library Amp Primer Mix, 96-well plate |
96-well Plate |
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For purchasing or general questions
For online ordering
*KAPA EvoPlus evaluation offer is valid for U.S. based customers only.