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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 signifcant improvements to yield, sensitivity, speed, target length, and the ability to amplify diffcult templates (e.g., AT- and GC-rich). The error rate of KAPA HiFi PCR Kits is 100X lower than wild-type Taq DNA polymerase.
Figure 3. 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.
Figure 4. 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.
Figure 5. KAPA HiFi DNA Polymerase exhibits robust performance on difficult templates. A panel of amplicons from human genomic DNA with a GC content ranging from 47% to 84% was used to compare the performance of KAPA HiFi HotStart and competitor kits. A polymerase with fusion technology in GC Buffer and a polymerase blend are only able to amplify targets up to 64% GC content. KAPA HiFi in GC Buffer achieves a 100% success rate with higher yields across all amplicons up to 84% GC content. Data on file.
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).
