KAPA HiFi DNA Polymerase (Hot Start formulation) is a novel, single-enzyme system that exhibits industry-leading fidelity and performance when compared with other proofreading polymerases and polymerase blends. KAPA HiFi has been engineered to have an increased affinity for DNA resulting in significant improvements to yield, sensitivity, speed, and the ability to amplify difficult templates. Strong proofreading activity results in an error rate 100X more accurate than Taq Polymerase.
KAPA HiFi HotStart DNA Polymerase is a novel B-family DNA polymerase, engineered to have increased affinity for DNA, without the need for accessory proteins or DNA binding domains. The intrinsic high processivity of the enzyme results in significant improvement in yield, speed and sensitivity when compared to wild-type B-family DNA polymerases. In addition, the ability to amplify long targets, as well as GC- and AT-rich targets, is significantly improved.
KAPA HiFi HotStart PCR Kits are designed for routine, high-fidelity PCR of a wide range of targets and fragment sizes. They offer error rates approximately 100 times lower than wild-type Taq DNA polymerase, and higher success rates and yields than achievable with wild-type B-family (proofreading) DNA polymerases. In addition, KAPA HiFi requires significantly shorter reaction times than wild-type B-family DNA polymerases.
KAPA HiFi HotStart DNA Polymerase has 5’g3’ polymerase and 3’g5’ exonuclease (proofreading) activity, but no 5’g3’ exonuclease activity. The strong 3’g5’ exonuclease activity results in superior accuracy during DNA amplification, lending to KAPA HiFi HotStart DNA Polymerase the lowest published error rate of all B-family DNA polymerases (1 error per 3.6 x 106 nucleotides incorporated). This fidelity is approximately 100 times higher than that of wild-type Taq DNA polymerase, and up to 10 times higher than that of other B-family DNA polymerases and polymerase blends.
DNA fragments generated with KAPA HiFi HotStart DNA Polymerase may be used for routine downstream analysis and applications, including restriction enzyme digestion, cloning and sequencing. PCR products generated with KAPA HiFi HotStart PCR Kits are blunt-ended, but may be 3’-dA-tailed for cloning into TA cloning vectors (see Important Parameters: TA cloning).
KAPA HiFi DNA Polymerase is supplied with two uniquelyformulated PCR buffers for optimal performance. Both buffers contain MgCl2 at a 1X concentration of 2 mM. The Fidelity Buffer is recommended for routine highfidelity PCR, while the GC Buffer is recommended for the amplification of GC-rich and other difficult targets. The additives in the GC Buffer result in a 2-fold decrease in fidelity when compared with the Fidelity Buffer.