KAPA HiFi PCR Kits

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.

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

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

Amplification of hgDNA targets up to 11 kb. Each target was amplified from a descending range of template hgDNA concentrations (50 ng to 0.5 ng per reaction). Reactions (25 µL each) were performed using standard 3-step cycling profiles (35 cycles): 20 sec denaturation, 15 sec annealing, and 30 sec/kb extension time. Total reaction time for the 11 kb amplicon was 3h 50 min. Data on file.

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

Applications

High-fidelity PCR
Amplification of DNA fragments for cloning and protein expression or genomic characterization

Site-directed mutagenesis
Next-generation sequencing library amplification

Kit Specifications and Contents / Storage

Store kits for 18 months at -20°C.

Kits include KAPA HiFi or KAPA HiFi HotStart DNA Polymerase (1 U/µL), Fidelity Buffer (5X), GC Buffer (5X), high-quality dNTPs (10 mM each) and MgCl2 (25 mM). Both buffers contain MgCl2 at a final concentration of 2 mM.

KAPA HiFi HotStart is also available in KAPA HiFi HotStart ReadyMix (2X), which contains DNA Polymerase, reaction buffer, dNTPs and MgCl2(at a final concentration of 2.5 mM).