Evolved for extreme fidelity, robustness, and low-bias amplification of difficult templates.

KAPA HiFi DNA Polymerase is a novel, single-enzyme system that exhibits industry-leading performance when compared with other high fidelity polymerases and polymerase blends. KAPA HiFi has been engineered to have an increased affinity for DNA without the need for accessory protein domains. The intrinsic high processivity of KAPA HiFi results in significant improvements to yield, sensitivity, speed, target length, and the ability to amplify difficult templates (e.g AT- and GC-rich). The error rate of KAPA HiFi PCR Kits is 100X lower than wild-type Taq DNA polymerase.

Amplifying NGS libraries? Go to our Library Amplification page.

For Research Use Only. Not for use in diagnostic procedures.

Product Highlights

The error rate of KAPA HiFi is 100x lower than Taq polymerase, 40x lower than polymerase blends such as Platinum® Taq High Fidelity, 3x lower than Pfu Ultra, and 2x lower than Phusion. Data on file.

Achieve the highest fidelity

Increased processivity, strong proofreading activity, and optimized buffer system result in superior accuracy
The error rate of KAPA HiFi is 100X lower than Taq, 40X lower than polymerase blends, 3X lower than Pfu Ultra and 2X lower than Phusion

Amplification of GC-rich DNA fragments. Amplification of different types of GC-rich DNA fragments using KAPA HiFi HotStart (top panel), a competitor engineered proofreading DNA polymerase containing a dsDNA-binding domain (hot start formulation, middle panel) and wild-type Pfu (bottom panel). Amplicon GC content increases from left (yellow) to right (red). Competitor reactions were performed in according to manufacturers’ instructions and were performed in GC Buffer (engineered competitor), or contained DMSO at a final concentration of 5% (Pfu). All reactions contained 25 ng human genomic DNA as template. Data on file.

Improve performance on GC- and AT-rich templates

Achieve higher success rates across targets up to 84% GC content
Higher yields when amplifying AT-rich templates

2 kb lambda phage target amplified from a 10-fold dilution series starting from 10 ng down to 10 fg using KAPA HiFi HotStart DNA Polymerase with Fidelity Buffer, fusion technology polymerase, and polymerase blend systems. All reactions (25 µL each) were performed using manufacturers’ protocols and buffers, with standard 3-step cycling profiles (35 cycles). Data on file.

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Amplify longer targets with greater sensitivity

High-fidelity PCR of long and complex genomic templates
Improved sensitivity
High speed allows significantly shorter reaction times for long range PCR

KAPA Library Amplification Kits

The gold standard for NGS library amplification.

KAPA Library Amplification Kits contain KAPA HiFi DNA Polymerase, a novel enzyme engineered for ultra-high fidelity and robustness using Kapa’s directed evolution technology platform.

KAPA HiFi has become the enzyme of choice for NGS library amplification due to its ability to amplify complex DNA populations with high fidelity, high efficiency, decreased PCR duplication rates and very low bias.* This results in lower duplication rates and improved coverage of GC- and AT rich regions, promoters, low complexity and other challenging regions in all NGS library construction workflows requiring library amplification.

In addition to the standard library amplification formulation, KAPA HiFi is available in a unique real-time formulation, for applications that demand precise control over library amplification. A uracil-tolerant variant, KAPA HiFi Uracil+ is also available for the high-efficiency, high-fidelity, low-bias amplification of libraries constructed from bisulfite-treated DNA.

KAPA Library Amplification Kits for Illumina® platforms now also include an optimally formulated Library Amplification Primer Mix.

*Data on file.

For Research Use Only. Not for use in diagnostic procedures.

Product Highlights

Coverage depth vs GC content for indexed Illumina TruSeq libraries prepared from 1 µg M. tuberculosis (65% GC) gDNA. Libraries amplified with KAPA HiFi HotStart or the Illumina TruSeq PCR Master Mix are compared to an equivalent unamplified library. Data was generated by paired-end sequencing (2 x 75 bp). After filtering and aligning read pairs to reference sequences, 250 000 read pairs were randomly sampled for each genome, and scatter plots of mean sequence coverage depth vs. GC content were generated by analyzing 250 bp windows. GC-rich sequences were under-represented in libraries amplified with the TruSeq PCR Master Mix. In contrast, library amplification with KAPA HiFi resulted in coverage distribution across the range of GC-content that is almost indistinguishable from that of the unamplified control. Data on file.

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Achieve the lowest amplification bias and duplication rates

Lower amplification bias result in improved representation of all library fragments and sequence regions
High-efficiency amplification leads to fewer amplification cycles and lower PCR duplicates
Less additional next-generation or Sanger sequencing needed to complete genomes

The first exons of many genes have a high GC content, and are therefore difficult to amplify. Regions of these exons are therefore typically underrepresented in whole exome sequence data. Pre-capture amplification with the evolved, low-bias KAPA HiFi DNA Polymerase results in significantly improved coverage of GC- and AT-rich regions, and other sequence elements that are difficult to amplify. Data courtesy of The Broad Institute. Data on file.

Improve coverage of GC- and AT-rich regions

Lower amplification bias improves coverage uniformity of GC- and AT-rich regions, promoters, low complexity and other challenging regions
Improved overall coverage and coverage uniformity observed on both Illumina and Ion Torrent™ sequencing platforms

Real-time, high fidelity amplification of multiple libraries with the KAPA Real-Time Library Amplification Kit. Twenty libraries, spanning a ~64-fold concentration range (equivalent to 6 cycles), were simultaneously amplified and terminated after 14 cycles. Fourteen of the twenty libraries fall within the targeted amplification range (the fluorescence range defined by the Real-Time Amplification Standards 1¬–3). The remaining six libraries could either be used as is, noting that they may be outside the optimal concentration range, or they could be re-amplified individually or in high- or low-concentration groups. Data on file.

Exercise precise control over library amplification

KAPA Real-Time Amplification Kits allow for library amplification to be observed in real time
Terminate amplification at the optimal point for individual samples
Optimize library amplification parameters for higher throughput workflows

Error rates of Taq DNA polymerase and polymerases typically used in NGS library amplification. The error rates of proofreading DNA polymerases such as Q5 (New England Biolabs), Phusion (Thermo Scientific) and the evolved KAPA HiFi DNA Polymerase is 50 – 100 times lower than that of Taq. The error rate of KAPA HiFi was confirmed by deep sequencing on the 454 platform, and is 1 error per 3.54 x 106 nucleotides incorporated. Data on file.

Amplify NGS libraries with industry-leading fidelity

KAPA HiFi was engineered using directed evolution to have enhanced proofreading (3′-5′ exonuclease) activity
Industry-leading fidelity confirmed by 454 sequencing