KAPA HiFi DNA Polymerase is a novel, single-enzyme system that exhibits industry-leading fidelity and performance when compared with other proofreading polymerases and polymerase blends. Strong proofreading activity results in an error rate 100X more accurate than Taq Polymerase.
What is Kapa PCR?
The KAPA Long Range PCR system is a blend of Taq DNA polymerase and an engineered archaeal (B-family) DNA polymerase possessing proofreading capability. This two-enzyme system is designed specifically to support long-range and/or sensitive PCR, and possesses 3 – 4X higher fidelity than Taq polymerase.
What should be annealing temperature in PCR?
The annealing temperature (typically between 48-72°C) is related to the melting temperature (Tm) of the primers and must be determined for each primer pair used in PCR. During the extension step (typically 68-72°C) the polymerase extends the primer to form a nascent DNA strand.
What happens if the annealing temperature is too high?
Annealing temperature was too high If the annealing temperature is too high, primers are unable to bind to the template. The rule of thumb is to use an annealing temperature that is 5°C lower than the Tm of the primer.
What is the difference between annealing and quenching?
After annealing, the grains are refined. the structure is adjusted, and the tissue defects are eliminated. Quenching causes the supercooled austenite to undergo martensite or bainite transformation. A martensite or bainite structure is obtained.
Why choose Kapa HiFi DNA polymerase?
The extreme processivity and robustness of KAPA HiFi DNA Polymerase offers the ability to perform high-fidelity PCR on long and complex templates with high speed and sensitivity.
What is Kapa HiFi Readymix?
KAPA HiFi HotStart ReadyMix (2X) is a ready-to-use cocktail containing all components for PCR, except primers and template. The 2X ReadyMix contains KAPA HiFi HotStart DNA Polymerase in proprietary reaction buffer, dNTPs (0.3 mM of each dNTP at 1X), MgCl (2.5 mM at 1X) and
How does Kapa HiFi perform on targets with high GC-content?
KAPA HiFi exhibits robust performance on targets with high GC-content. A panel of human amplicons with GC contents ranging from 66 – 84% was used to compare the performance of KAPA HiFi with that of an engineered polymerase with fusion technology and wild-type Pfu. KAPA HiFi achieved a 100% success rate with higher yields up to 84% GC content.
What is the error rate of Kapa HiFi?
The error rate of KAPA HiFi is 100X lower than that of TaqPolymerase, 40X lower than that of polymerase blends such as Platinum®Taq High Fidelity, 3X lower than that of PfuUltra™, and 2X lower than that of Phusion™. Fidelity data for Taq, PfuUltra™ and Phusion™ are based on published error rates.
